Nucleic acid molecules associated with the methionine synthesis and degradation pathways

ABSTRACT

The present invention is in the field of plant biochemistry. More specifically the invention relates to nucleic acid sequences from plant cells, in particular, DNA sequences from maize and soybean plants associated with the methionine pathway. The invention encompasses nucleic acid molecules that encode proteins and fragments of proteins. In addition, the invention also encompasses proteins and fragments of proteins so encoded and antibodies capable of binding these proteins or fragments. The invention also relates to methods of using the nucleic acid molecules, proteins and fragments of proteins and antibodies, for example for genome mapping, gene identification and analysis, plant breeding, preparation of constructs for use in plant gene expression and transgenic plants.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No. 09/198,779 filed Nov. 24, 1998, now abandoned which claims the benefit under 35 U.S.C. ±119(e) of U.S. Applications No. 60/067,000 filed Nov. 24, 1997, No. 60/066,873 filed Nov. 25, 1997, No. 60/069,472 filed Dec. 9, 1997, No. 60/074,201 filed Feb. 10, 1998, No. 60/074,282 filed Feb. 10, 1998, No. 60/074,280 filed Feb. 10, 1998, No. 60/074,281 filed Feb. 10, 1998, No. 60/074,566 filed Feb. 12, 1998, No. 60/074,567 filed Feb. 12, 1998, No. 60/074,565 filed Feb. 12, 1998, No. 60/075,462 filed Feb. 19, 1998, No. 60/074,789 filed Feb. 19, 1998, No. 60/075,459 filed Feb. 19, 1998, No. 60/075,461 filed Feb. 19, 1998, No. 60/075,464 filed Feb. 19, 1998, No. 60/075,460 filed Feb. 19, 1998, No. 60/075,463 filed Feb. 19, 1998, No. 60/077,231 filed Mar. 9, 1998, No. 60/077,229 filed Mar. 9, 1998, No. 60/077,230 filed Mar. 9, 1998, No. 60/078,031 filed Mar. 16, 1998, No. 60/078,368 filed Mar. 18, 1998, No. 60/080,844 filed Apr. 7, 1998, No. 60/083,067 filed Apr. 27, 1998, No. 60/083,386 filed Apr. 29, 1998, No. 60/083,387 filed Apr. 29, 1998, No. 60/083,388 filed Apr. 29, 1998, No. 60/083,389 filed Apr. 29, 1998, No. 60/084,684 filed May 8, 1998, No. 60/085,245 filed May 13, 1998, No. 60/085,224 filed May 13, 1998, No. 60/085,223 filed May 13, 1998, No. 60/085,222 filed May 13, 1998, No. 60/086,186 filed May 21, 1998, No. 60/086,339 filed May 21, 1998, No. 60/086,187 filed May 21, 1998, No. 60/086,185 filed May 21, 1998, No. 60/086,184 filed May 21, 1998, No. 60/086,183 filed May 21, 1998, No. 60/086,188 filed May 21, 1998, No. 60/089,524 filed Jun. 16, 1998, No. 60/089,810 filed Jun. 18, 1998, No. 60/089,814 filed Jun. 18, 1998, No. 60/091,247 filed Jun. 30, 1998, No. 60/092,036 filed Jul. 8, 1998, No. 60/099,667 filed Sep. 9, 1998, No. 60/099,668 filed Sep. 9, 1998, No. 60/099,670 filed Sep. 9, 1998, No. 60/099,697 filed Sep. 9, 1998, No. 60/100,674 filed Sep. 16, 1998, No. 60/100,673 filed Sep. 16, 1998, No. 60/100,672 filed Sep. 16, 1998, No. 60/101,132 filed Sep. 21, 1998, No. 60/101,130 filed Sep. 21, 1998, No. 60/101,508 filed Sep. 21, 1998, No. 60/101,344 filed Sep. 22, 1998, No. 60/101,347 filed Sep. 22, 1998, No. 60/101,343 filed Sep. 22, 1998, No. 60/104,126 filed Oct. 13, 1998, No. 60/104,128 filed Oct. 13, 1998, No. 60/104,127 filed Oct. 13, 1998, No. 60/104,124 filed Oct. 13, 1998, No. 60/109,018 filed Nov. 18, 1998 and No. 60/108,996 filed Nov. 18, 1998 hereby incorporated by reference herein in their entirety.

Two copies of the sequence listing (Copy 1 and Copy 2) and a computer readable form of the sequence listing, all on CD-ROMs, each filed containing the file named “16517328_seq.txt” which is 1,821,718 bytes in size (measured in MS-DOS) and was created on Oct. 7, 2004, are herein incorporated by reference.

FIELD OF THE INVENTION

The present invention is in the field of plant biochemistry. More specifically the invention relates to nucleic acid sequences from plant cells, in particular, DNA sequences from maize and soybean plants associated with the methionine pathway. The invention encompasses nucleic acid molecules that encode proteins and fragments of proteins. In addition, the invention also encompasses proteins and fragments of proteins so encoded and antibodies capable of binding these proteins or fragments. The invention also relates to methods of using the nucleic acid molecules, proteins and fragments of proteins and antibodies, for example for genome mapping, gene identification and analysis, plant breeding, preparation of constructs for use in plant gene expression and transgenic plants.

BACKGROUND OF THE INVENTION I. Methoinine Synthesis Pathway

The amino acid, L-methionine, is synthesized in higher plants via a pathway that starts with L-aspartate. This pathway has been studied (Azevedo et al., Phytochemistry 46:395-419 (1997), the entirety of which is herein incorporated by reference). L-methionine is one of four so-called aspartate-derived amino acids (along with L-lysine, L-threonine and L-isoleucine) (Miflin et al., In: Nitrogen Assimilation in Plants, Hewitt et al., (eds.), Academic Press, New York, 335 (1997); Bryan, In: The Biochemistry of Plants, Miflin (ed.), Academic Press, New York, 403 (1980); Lea et al., In: The Chemistry and Biochemistry of Amino Acids, Barrett et al., (eds.), London, 5:197 (1985); Bryan, In: The Biochemistry of Plants, Miflin et al., (eds.), Academic Press, San Diego, 16:161 (1990), all of which are herein incorporated by reference in their entirety).

The methionine-specific part of the aspartate pathway includes the following enzymes: aspartate kinase (EC 2.7.2.4), aspartate-semialdehyde dehydrogenase (EC 1.2.1.11), homoserine dehydrogenase (EC 1.1.1.3), homoserine kinase (EC 2.7.1.39), cystathionine γ-synthase (EC 4.2.99.9), cystathionine β-lyase (EC 4.4.1.8) and methionine synthase (EC 2.1.1.14).

Aspartate kinase catalyzes the first reaction of the pathway in which aspartate is converted to β-aspartyl phosphate. This enzyme has been isolated and characterized from plant sources including maize, barley, carrot, pea and soybean. These studies have revealed that there are multiple isoenzymes of aspartate kinase and the isoenzymes differ with respect to both feedback inhibition sensitivity and expression profile (tissue and developmental stage). Feedback inhibition is mediated by lysine and threonine. Transgenic plants which express an unregulated aspartate kinase have demonstrated increased flux through the aspartate pathway. Pathway regulation is reported to be exerted, at least in part, via control of this enzyme's activity.

Aspartate semialdehyde dehydrogenase catalyses the second pathway reaction and converts β-aspartyl phosphate to aspartate semialdehyde via an NADPH-dependent reaction. Gengenbach et al., Crop Science 18:472-476 (1978), the entirety of which is herein incorporated by reference, report the isolation of aspartate semialdehyde dehydrogenase from maize suspension culture cells. These suspension cultures did not exhibit feedback inhibition of the enzyme in the presence of aspartate-derived amino acids, with the exception of methionine, for which some feedback sensitivity was observed. Aspartate semialdehyde dehydrogenase enzyme activity has been reported in maize shoot, maize root and maize kernel (Gengenbach et al., Crop Science 18:472-476 (1978)).

Homoserine dehydrogenase catalyzes the next step of the pathway in which homoserine is generated from aspartate semialdehyde in a reaction requiring NADH or NADPH. Homoserine dehydrogenase enzyme has been studied in higher plants and multiple isoenzyme forms have been reported (Bryan et al., Biochemistry and Biophysics Research Communications 41:1211-1217 (1970); Gengenbach et al., Crop Science 18:472-476 (1978); Dotson et al., Plant Physiology 91:1602-1608 (1989); Dotson et al., Plant Physiology 93:98-104 (1989); Azevedo et al., Phytochemistry 31:3725-3730 (1992); Azevedo et al., Phytochemistry 31:3731-3734 (1992); Brennecke et al., Phytochemistry 41:707 (1996); Aarnes, Plant Science Letters 9:137-145 (1977); Bright et al., Biochemical Genetics 200:229-243 (1982); Aruda et al., Plant Physiology 76:442-446 (1984); Lea et al., In: Barley: Genetics, Molecular Biology and Biotechnology Shewrey (ed.), CAB International, Oxford 181 (1992); Davies et al., Plant Science Letters 9:323-332 (1977); Davies et al., Plant Physiology 62:536-541 (1978); Matthews et al., Zeitschrift für Naturforschung, Section Bioscience 34:1177-1185 (1979); Relton et al., Biochimica et Biophysica Acta 953:48-60 (1988); Aarnes et al., Phytochemistry 13:2717-2724 (1974); Lea et al., FEBS Letters 98:165-168 (1979); Matthews et al., Canadian Journal of Botany 57:299-304 (1979), all of which references are incorporated herein in their entirety). The isoenzymes have been found to differ with respect to sensitivity to threonine-mediated feedback inhibition, with both sensitive and insensitive forms being isolated from maize suspension cultures and seedlings (Miflin et al., In: Nitrogen Assimilation of Plants, Hewitt et al., (eds.), Academic Press, New York, 335 (1997); Bryan, In: The Biochemistry of Plants, Miflin (ed.), Academic Press, New York, 5:403 (1980)).

There is evidence that plants also possess a bifunctional enzyme with both aspartate kinase and homoserine dehydrogenase activities (Lea et al., In: The Chemistry and Biochemistry of Amino Acids, Barrett et al. (eds), London, 5:197 (1985), the entirety of which is herein incorporated by reference; Bryan, In: The Biochemistry of Plants, Miflin (ed.), Academic Press, New York, 5:161 (1990), the entirety of which is herein incorporated by reference). Clones of these bifunctional enzymes have been isolated from Arabidopsis thaliana (Giovanelli et al., In: The Biochemistry of Plants, Miflin (ed.), Academic Press, New York 453 (1990), the entirety of which is herein incorporated by reference) carrot (Giovanelli et al., Plant Physiology 90:1584-1599 (1989), the entirety of which is herein incorporated by reference), maize (Singh et al., Amino Acids 7:165-168 (1994), the entirety of which is herein incorporated by reference) and soybean (Matthews et al., In: Biosynthesis and Molecular Regulation of Amino Acids in Plants, p 294, Singh et al. (eds.), American Society of Plant Physiologists, Rockville, Md. (1992), the entirety of which is herein incorporated by reference).

The next reported enzymatic step leading to methionine biosynthesis in higher plants is the final common reaction shared by other amino acid end products (threonine and isoleucine). The reaction is catalyzed by homoserine kinase and it generates O-phosphohomoserine from homoserine, with ATP serving as the phosphate donor. Exceptions are Pisum sativum and Lathyrus sitivus which synthesize O-acetylhomoserine and O-oxalylhomoserine, respectively (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532 (1997), the entirety of which is herein incorporated by reference). Enteric bacteria use O-succinylhomoserine, while several gram-positive bacteria, yeasts and fungi use O-acetylhomoserine (formed using homoserine O-acetyltransferase (EC 2.3.1.31) (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532 (1997)). Homoserine kinase has been reported from multiple higher plant sources (Galili, The Plant Cell 7:899-906 (1995), the entirety of which is herein incorporated by reference; Rees et al., Biochemical Journal 309:999-1107 (1995), the entirety of which is herein incorporated by reference; Bryan et al., Biochemistry and Biophysics Research Communications 41:1211-1217 (1970), the entirety of which is herein incorporated by reference; Gengenbach et al., Crop Science 18:472-476 (1978), Dotson et al., Plant Physiology 91:1602-1608 (1989), the entirety of which is herein incorporated by reference; Dotson et al., Plant Physiology 93:98-104 (1989), the entirety of which is herein incorporated by reference). Homoserine kinase isolated from barley and wheat has not been reported to exhibit aspartate-derived amino acid feedback inhibition (Gengenbach et al., Crop Science 18:472-476 (1978); Dotson et al., Plant Physiology 93:98-104 (1989)). It has been reported that homoserine kinase exhibits feedback regulation in the dicots, pea (Rees et al., Biochemical Journal 309:999-1007 (1995), the entirety of which is herein incorporated by reference) and radish (Bryan et al., Biochemistry and Biophysics Research Communications 41:1211-1217 (1970)). Bacterial and yeast homologues have been reported (Azevedo et al., Phytochemistry 31:3725-3730 (1992); Azevedo et al., Phytochemistry 31:3731-3734 (1992); Brennecke et al., Phytochemistry 41:707 (1996); Aarnes, Plant Science Letters 9:137-145 (1977)).

Sulfur, in yeast, is incorporated into O-acetylhomoserine resulting in homocysteine. This reaction is catalyzed by the O-acetylhomoserine sulfhydrylase (EC 4.2.99.10) (also known as O-acethomoserine (thiol)-lyase). O-acetylhomoserine sulfhydrylase has been reported to be a homotetramer with a molecular weight of 200,000. O-acetylhomoserine sulfhydrylase has also been reported to bind four molecules of pyridoxal phosphate (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532 (1997)).

In higher plants, the sulfur atom from cysteine and the carbon backbone derived from aspartate used to synthesize methionine are reported to be catalyzed by pyridoxal 5′-phosphate (PLP) dependent enzymes (Ravanel et al., Proc. Natl. Acad. Sci. (U.S.A.) 95:7805-7812 (1998), the entirety of which is herein incorporated by reference). The amino acid composition of the O-acetylhomoserine sulfhydrylase has also been reported to share sequence similarities to the E. coli cystathionine γ-synthase and cystathionine β-lyase and cystathionine γ-lyase from Saccharomyces cervisiae and rats. All of these enzymes thus appear to belong to one protein family, whose members have evolved from an ancestral pyridoxal phosphate enzyme (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532 (1997)).

In yeast, the synthesis of cysteine from homocysteine has been reported to require two successive steps, β addition and γ elimination. Cystathionine β-synthase (EC 4.2.1.22) has been reported to catalyze the first reaction where homocysteine and serine yield cystathionine. In S. cervisiae, cystathionine β-synthase is encoded by STR4. STR4 encodes a polypeptide of 506 residues which shows extensive sequence similarity to its functional analog in rats. The rat analog has been reported to contain an additional amino-terminal extension of 60 residues. Moreover, the two enzymes have been reported to be closely related to the cysteine synthase from enteric bacteria and plants (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532 (1997)).

Cystathionine γ-lyase (EC 4.4.1.1) catalyzes the γ cleavage of cystationine in yeast, the second reported step of the biosynthesis of cysteine from homocysteine. Cystathionine γ-lyase has been reported to have a molecular weight of about 194,000 kd. In S. cerviseae, cystathionine γ-lyase is encoded by STR1. A mutation in the S. cerviseae cystathionine γ-lyase gene leads to a nutritional requirement for cysteine or glutathione. The yeast cystathionine γ-lyase belongs to a protein family which includes a functional analog in rats, a Met25p from yeast and cystathionin β-lyase and cystathionin γ-synthase from E. coli (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532 (1997)).

Cystathionine γ-synthase (also known as O-succinylhomoserine (thio)-lyase, E.C. 4.2.99.9) catalyzes the first reported reaction which is unique to methionine biosynthesis, thereby committing aspartate pathway flux toward this amino acid. In this reaction, O-phosphohomoserine and cysteine serve as substrates for the production of cystathionine. Cystathionine γ-synthase has not been reported to be regulated by aspartate-derived amino acids feedback inhibition (Bright et al., Biochemical Genetics 20:229-243 (1982); Arruda et al., Plant Physiology 76:442-446 (1984)). Cystathionine γ-synthase has however, been reported to be sensitive to product inhibition by orthophosphate (Lea et al., Barley: Genetics, Molecular Biology and Biotechnology, Shewrey (ed.), CAB International, Oxford, 181 (1992); Davies et al., Plant Science Letters 9:323-332 (1977)). Cloned cystathionine γ-synthase have been reported from Arabidopsis thaliana (Davies et al., Plant Physiology 62:536-541 (1978)). It has been reported that methionine levels are modulated via regulation of cystathionine-synthase (Matthews et al., Zeitschrift für Naturforschung, Section Bioscience 34:1177-1185 (1979-2724 (1974); Lea et al., FEBS Letters 98:165 (1979), all of which references are incorporated herein in their entirety).

Cystathionine β-lyase catalyzes the next reaction in the biosynthesis of methionine. This reaction generates homocysteine, pyruvate and ammonia from the enzymatic decomposition of cystathionine. Evidence for isoenzymes which differ with respect to cellular localization have been reported for barley (Matthews et al., Canadian Journal of Botany 57:299-304 (1979)) and spinach (Rognes et al., Nature 287:357-359 (1980), the entirety of which is herein incorporated by reference).

De novo synthesis of methionine from homocysteine uses a methyl group which originates from single-carbon metabolism. In this metabolism, derivatives of tetrahydrofolate transfer one-carbon groups at the oxidation levels of methanol, formaldehyde and formate to acceptor molecules. Single-carbon derivatives of tetrahydrofolate are required for the biosynthesis of methionine, purine nucleotides and thymidylate as well as for the synthesis of N-formylmethionine in the mitochondrion. S. cerevisiae possesses two complete sets of folate interconversion enzymes, one located in the cytosol (methionyl-tRNA synthetase, EC 6.1.1.10) and the other located in the mitochondrion (methionyl t-RNA synthetase, EC 6.1.1.10) (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532 (1997)) and in plants including the chloroplast (Menand et al., Proc. Natl. Acad. Sci. (U.S.A.), 95:11014-11019 (1998), the entirety of which is herein incorporated by reference).

Methionine synthase generates methionine from homocysteine by a methylation reaction and thus represents the final step of the methionine biosynthetic pathway. Methionine synthase is also sometimes referred to as 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase. N-methyltetrahydrofolate serves as the methyl donor in this reaction, which occurs in the absence of cobalamin (Giovanelli et al., Plant Physiology 90:1577-1583 (1989), the entirety of which is herein incorporated by reference; Green et al., Crop Science 14:827-830 (1974), the entirety of which is herein incorporated by reference).

II. Methoinine Degradation Pathway

Plants contain a pathway for the degradation of L-methionine. This degradation pathway includes the following enzymes: methionine adenosyltransferase (EC 2.5.1.6), methionine S-methyltransferase (EC 2.1.1.12), adenosylmethionine hydrolase (EC 3.3.1.2), homocysteine S-methyltransferase (EC 2.1.1.10) and S-adenosyl-methionine decarboxylase (EC 4.1.1.50).

The reported first step in the catabolism of methionine is the ATP-dependent conversion to S-adenosylmethionine (AdoMet), which is catalyzed by the enzyme methionine adenosyltransferase, also known as S-adenosylmethionine synthetase. Methionine adenosyltransferase enzyme has been characterized from several plant sources (Aarnes, Plant Science Letters 10:381 (1977), the entirety of which is herein incorporated by reference; Mathur et al., Biochimia and Biophysica Acta 1078:161-170 (1991), the entirety of which is herein incorporated by reference; Kim et al., Journal of Biochemical and Molecular Biology 28:100 (1995), the entirety of which is herein incorporated by reference) and nucleic acid molecules (genomic and cDNA) have also been obtained from a variety of sources (Izhaki et al., Plant Physiology 108:841-842 (1995), the entirety of which is herein incorporated by reference; Espartero et al., Molecular Biology Plant 25:217-237 (1994), the entirety of which is herein incorporated by reference). Regulation of methionine adenosyltransferase activity has been observed for the enzyme from Glycine max (soybean). In Glycine max, methionine adenosyltransferase was reportedly inhibited by S-adenosylmethionine (Kim et al., Journal of Biochemical and Molecular Biology 28:100 (1995). Studies have also reported that the levels of methionine adenosyltransferase appear to fluctuate in response to hormonal or environmental conditions such as gibberellic acid (Mathur et al., Biochimica and Biophysica Acta 1162:289-290 (1993), the entirety of which is herein incorporated by reference; Mathur et al., Biochimica and Biophysica Acta 1137:338-348 (1992), the entirety of which is herein incorporated by reference), salt stress (Espartero et al., Molecular Biology Plant 25:217-227 (1994) the entirety of which is herein incorporated by reference) and wounding (Kim et al., Plant Cell Reports 13:340 (1994), the entirety of which is herein incorporated by reference). It has also been reported that methionine adenosyltransferase may play a role in the lignification process (Peleman et al., Plant Cell 1:81 (1989), the entirety of which is herein incorporated by reference).

AdoMet is further catabolized by several enzymes and has been reported to serve a variety of metabolic functions including that of a methyl donor (Cossins, The Biochemistry of Plants 11:317 Devis (ed.), Academic Press, San Diego (1987), the entirety of which is herein incorporated by reference) that of a precursor for polyamine biosynthesis (Tiburico et al., The Biochemistry of Plants 16:283 (1990), the entirety of which is herein incorporated by reference) and that of a precursor for ethylene biosynthesis (Kende, Plant Physiology 91:1-4 (1989), the entirety of which is herein incorporated by reference; Flurh et al., Critical Review of Plant Science 15:479 (1996), the entirety of which is herein incorporated by reference). In each case, enzymes are present to regenerate methionine from the sulfur-containing backbone resulting in no net loss of methionine.

An enzyme involved in AdoMet catabolism is adenosylmethionine hydrolase (EC 3.3.1.2) which converts AdoMet to methylthioadenosine and L-homoserine. L-homoserine is further metabolized during the biosynthesis of polyamines and ethylene and methylthioadenosine is recycled to methionine. In yeast, a form of adenosylmethioninehydrolase (EC 3.1.1.1) has been reported (on the Worldwide web at ncbi.nlm.nih.gov/htbin-post/Entrez/query (1998)).

Another enzyme for which AdoMet is a substrate for is homocysteine S-methyltransferase. Homocysteine S-methyltransferase catalyzes the combination of AdoMet, with L-homocysteine to produce both S-adenosyl-L-homocysteine and L-methionine. Another enzyme has been described which generates S-adenosyl-L-homocysteine from AdoMet. This enzyme is called methionine S-methyltransferase and it catalyzes the reaction in which S-adenosyl-L-homocysteine reacts with L-methionine to generate S-adenosyl-L-homocysteine and S-methyl-L-methionine. AdoMet can also be decarboxylated by adenosyl methionine decarboxylase, which generates (5-deoxy-5-adenosyl)(3-aminopropyl) methylsulfonium salt.

III. Expressed Sequence Tag Nucleic Acid Molecules

Expressed sequence tags, or ESTs are randomly sequenced members of a cDNA library (or complementary DNA) (McCombie et al., Nature Genetics 1:124-130 (1992); Kurata et al., Nature Genetics 8:365-372 (1994); Okubo et al., Nature Genetics 2:173-179 (1992), all of which references are incorporated herein in their entirety). The randomly selected clones comprise insets that can represent a copy of up to the full length of a mRNA transcript.

Using conventional methodologies, cDNA libraries can be constructed from the mRNA (messenger RNA) of a given tissue or organism using poly dT primers and reverse transcriptase (Efstratiadis et al., Cell 7:279-3680 (1976), the entirety of which is herein incorporated by reference; Higuchi et al., Proc. Natl. Acad. Sci. (U.S.A.) 73:3146-3150 (1976), the entirety of which is herein incorporated by reference; Maniatis et al., Cell 8:163-182 (1976) the entirety of which is herein incorporated by reference; Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety of which is herein incorporated by reference; Okayama et al., Mol. Cell. Biol. 2:161-170 (1982), the entirety of which is herein incorporated by reference; Gubler et al., Gene 25:263-269 (1983), the entirety of which is herein incorporated by reference).

Several methods may be employed to obtain full-length cDNA constructs. For example, terminal transferase can be used to add homopolymeric tails of dC residues to the free 3′ hydroxyl groups (Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety of which is herein incorporated by reference). This tail can then be hybridized by a poly dG oligo which can act as a primer for the synthesis of full length second strand cDNA. Okayama and Berg, Mol. Cell. Biol. 2:161-170 (1982), the entirety of which is herein incorporated by reference, report a method for obtaining full length cDNA constructs. This method has been simplified by using synthetic primer-adapters that have both homopolymeric tails for priming the synthesis of the first and second strands and restriction sites for cloning into plasmids (Coleclough et al., Gene 34:305-314 (1985), the entirety of which is herein incorporated by reference) and bacteriophage vectors (Krawinkel et al., Nucleic Acids Res. 14:1913 (1986), the entirety of which is herein incorporated by reference; Han et al., Nucleic Acids Res. 15:6304 (1987), the entirety of which is herein incorporated by reference).

These strategies have been coupled with additional strategies for isolating rare mRNA populations. For example, a typical mammalian cell contains between 10,000 and 30,000 different mRNA sequences (Davidson, Gene Activity in Early Development, 2nd ed., Academic Press, New York (1976), the entirety of which is herein incorporated by reference). The number of clones required to achieve a given probability that a low-abundance mRNA will be present in a cDNA library is N=(ln(1−P))/(ln(1−1/n)) where N is the number of clones required, P is the probability desired and 1/n is the fractional proportion of the total mRNA that is represented by a single rare mRNA (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press (1989), the entirety of which is herein incorporated by reference).

A method to enrich preparations of mRNA for sequences of interest is to fractionate by size. One such method is to fractionate by electrophoresis through an agarose gel (Pennica et al., Nature 301:214-221 (1983), the entirety of which is herein incorporated by reference). Another such method employs sucrose gradient centrifugation in the presence of an agent, such as methylmercuric hydroxide, that denatures secondary structure in RNA (Schweinfest et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:4997-5000 (1982), the entirety of which is herein incorporated by reference).

A frequently adopted method is to construct equalized or normalized cDNA libraries (Ko, Nucleic Acids Res. 18:5705-5711 (1990), the entirety of which is herein incorporated by reference; Patanjali et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:1943-1947 (1991), the entirety of which is herein incorporated by reference). Typically, the cDNA population is normalized by subtractive hybridization (Schmid et al., J. Neurochem. 48:307-312 (1987), the entirety of which is herein incorporated by reference; Fargnoli et al., Anal. Biochem. 187:364-373 (1990), the entirety of which is herein incorporated by reference; Travis et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:1696-1700 (1988), the entirety of which is herein incorporated by reference; Kato, Eur. J. Neurosci. 2:704-711 (1990); and Schweinfest et al., Genet. Anal. Tech. Appl. 7:64-70 (1990), the entirety of which is herein incorporated by reference). Subtraction represents another method for reducing the population of certain sequences in the cDNA library (Swaroop et al., Nucleic Acids Res. 19:1954 (1991), the entirety of which is herein incorporated by reference).

ESTs can be sequenced by a number of methods. Two basic methods may be used for DNA sequencing, the chain termination method of Sanger et al., Proc. Natl. Acad. Sci. (U.S.A.) 74:5463-5467 (1977), the entirety of which is herein incorporated by reference and the chemical degradation method of Maxam and Gilbert, Proc. Nat. Acad. Sci. (U.S.A.) 74:560-564 (1977), the entirety of which is herein incorporated by reference. Automation and advances in technology such as the replacement of radioisotopes with fluorescence-based sequencing have reduced the effort required to sequence DNA (Craxton, Methods 2:20-26 (1991), the entirety of which is herein incorporated by reference; Ju et al., Proc. Natl. Acad. Sci. (U.S.A.) 92:4347-4351 (1995), the entirety of which is herein incorporated by reference; Tabor and Richardson, Proc. Natl. Acad. Sci. (U.S.A.) 92:6339-6343 (1995), the entirety of which is herein incorporated by reference). Automated sequencers are available from, for example, Pharmacia Biotech, Inc., Piscataway, N.J. (Pharmacia ALF), LI-COR, Inc., Lincoln, Nebr. (LI-COR 4,000) and Millipore, Bedford, Mass. (Millipore BaseStation).

In addition, advances in capillary gel electrophoresis have also reduced the effort required to sequence DNA and such advances provide a rapid high resolution approach for sequencing DNA samples (Swerdlow and Gesteland, Nucleic Acids Res. 18:1415-1419 (1990); Smith, Nature 349:812-813 (1991); Luckey et al., Methods Enzymol. 218:154-172 (1993); Lu et al., J. Chromatog. A. 680:497-501 (1994); Carson et al., Anal. Chem. 65:3219-3226 (1993); Huang et al., Anal. Chem. 64:2149-2154 (1992); Kheterpal et al., Electrophoresis 17:1852-1859 (1996); Quesada and Zhang, Electrophoresis 17:1841-1851 (1996); Baba, Yakugaku Zasshi 117:265-281 (1997), all of which are herein incorporated by reference in their entirety).

ESTs longer than 150 nucleotides have been found to be useful for similarity searches and mapping (Adams et al., Science 252:1651-1656 (1991), herein incorporated by reference). ESTs, which can represent copies of up to the full length transcript, may be partially or completely sequenced. Between 150-450 nucleotides of sequence information is usually generated as this is the length of sequence information that is routinely and reliably produced using single run sequence data. Typically, only single run sequence data is obtained from the cDNA library (Adams et al., Science 252:1651-1656 (1991). Automated single run sequencing typically results in an approximately 2-3% error or base ambiguity rate (Boguski et al., Nature Genetics 4:332-333 (1993), the entirety of which is herein incorporated by reference).

EST databases have been constructed or partially constructed from, for example, C. elegans (McCombrie et al., Nature Genetics 1: 124-131 (1992)), human liver cell line HepG2 (Okubo et al., Nature Genetics 2:173-179 (1992)), human brain RNA (Adams et al., Science 252:1651-1656 (1991); Adams et al., Nature 355:632-635 (1992)), Arabidopsis, (Newman et al., Plant Physiol. 106:1241-1255 (1994)); and rice (Kurata et al., Nature Genetics 8:365-372 (1994)).

IV. Sequence Comparisons

A characteristic feature of a DNA sequence is that it can be compared with other DNA sequences. Sequence comparisons can be undertaken by determining the similarity of the test or query sequence with sequences in publicly available or proprietary databases (“similarity analysis”) or by searching for certain motifs (“intrinsic sequence analysis”) (e.g. cis elements) (Coulson, Trends in Biotechnology 12:76-80 (1994), the entirety of which is herein incorporated by reference); Birren et al., Genome Analysis 1: Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 543-559 (1997), the entirety of which is herein incorporated by reference).

Similarity analysis includes database search and alignment. Examples of public databases include the DNA Database of Japan (DDBJ) (on the Worldwide web at ddbj.nig.ac.jp/; GENBANK (on the Worldwide web at ncbi.nlm.nih.gov/Web/SearchlIndex.htlm); and the European Molecular Biology Laboratory Nucleic Acid Sequence Database (EMBL) (on the Worldwide web at ebi.ac.uk/ebi_docs/embl_db/embl_db.html. Other appropriate databases include dbEST (on the Worldwide web at ncbi.nlm.nih.gov/dbEST/index.html, SwissProt (on the Worldwide web at ebi.ac.uk/ebi_docs/swisprot_db/swisshome.html, PIR (on the Worldwide web at nbrt.georgetown.edu/pir/ and The Institute for Genome Research (on the Worldwide web at tigr.org/tdb/tdb.html).

A number of different search algorithms have been developed, one example of which are the suite of programs referred to as BLAST programs. There are five implementations of BLAST, three designed for nucleotide sequences queries (BLASTN, BLASTX and TBLASTX) and two designed for protein sequence queries (BLASTP and TBLASTN) (Coulson, Trends in Biotechnology 12:76-80 (1994); Birren et al., Genome Analysis 1, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 543-559 (1997)).

BLASTN takes a nucleotide sequence (the query sequence) and its reverse complement and searches them against a nucleotide sequence database. BLASTN was designed for speed, not maximum sensitivity and may not find distantly related coding sequences. BLASTX takes a nucleotide sequence, translates it in three forward reading frames and three reverse complement reading frames and then compares the six translations against a protein sequence database. BLASTX is useful for sensitive analysis of preliminary (single-pass) sequence data and is tolerant of sequencing errors (Gish and States, Nature Genetics 3:266-272 (1993), the entirety of which is herein incorporated by reference). BLASTN and BLASTX may be used in concert for analyzing EST data (Coulson, Trends in Biotechnology 12:76-80 (1994); Birren et al., Genome Analysis 1:543-559 (1997)).

Given a coding nucleotide sequence and the protein it encodes, it is often preferable to use the protein as the query sequence to search a database because of the greatly increased sensitivity to detect more subtle relationships. This is due to the larger alphabet of proteins (20 amino acids) compared with the alphabet of nucleic acid sequences (4 bases), where it is far easier to obtain a match by chance. In addition, with nucleotide alignments, only a match (positive score) or a mismatch (negative score) is obtained, but with proteins, the presence of conservative amino acid substitutions can be taken into account. Here, a mismatch may yield a positive score if the non-identical residue has physical/chemical properties similar to the one it replaced. Various scoring matrices are used to supply the substitution scores of all possible amino acid pairs. A general purpose scoring system is the BLOSUM62 matrix (Henikoff and Henikoff, Proteins 17:49-61 (1993), the entirety of which is herein incorporated by reference), which is currently the default choice for BLAST programs. BLOSUM62 is tailored for alignments of moderately diverged sequences and thus may not yield the best results under all conditions. Altschul, J. Mol. Biol. 36:290-300 (1993), the entirety of which is herein incorporated by reference, describes a combination of three matrices to cover all contingencies. This may improve sensitivity, but at the expense of slower searches. In practice, a single BLOSUM62 matrix is often used but others (PAM40 and PAM250) may be attempted when additional analysis is necessary. Low PAM matrices are directed at detecting very strong but localized sequence similarities, whereas high PAM matrices are directed at detecting long but weak alignments between very distantly related sequences.

Homologues in other organisms are available that can be used for comparative sequence analysis. Multiple alignments are performed to study similarities and differences in a group of related sequences. CLUSTAL W is a multiple sequence alignment package that performs progressive multiple sequence alignments based on the method of Feng and Doolittle, J. Mol. Evol. 25:351-360 (1987), the entirety of which is herein incorporated by reference. Each pair of sequences is aligned and the distance between each pair is calculated; from this distance matrix, a guide tree is calculated and all of the sequences are progressively aligned based on this tree. A feature of the program is its sensitivity to the effect of gaps on the alignment; gap penalties are varied to encourage the insertion of gaps in probable loop regions instead of in the middle of structured regions. Users can specify gap penalties, choose between a number of scoring matrices, or supply their own scoring matrix for both pairwise alignments and multiple alignments. CLUSTAL W for UNIX and VMS systems is available at: ftp.ebi.ac.uk. Another program is MACAW (Schuler et al., Proteins Struct. Func. Genet. 9:180-190 (1991), the entirety of which is herein incorporated by reference, for which both Macintosh and Microsoft Windows versions are available. MACAW uses a graphical interface, provides a choice of several alignment algorithms and is available by anonymous ftp at: ncbi.nlm.nih.gov (directory/pub/macaw).

Sequence motifs are derived from multiple alignments and can be used to examine individual sequences or an entire database for subtle patterns. With motifs, it is sometimes possible to detect distant relationships that may not be demonstrable based on comparisons of primary sequences alone. Currently, the largest collection of sequence motifs in the world is PROSITE (Bairoch and Bucher, Nucleic Acid Research 22:3583-3589 (1994), the entirety of which is herein incorporated by reference). PROSITE may be accessed via either the ExPASy server on the World Wide Web or anonymous ftp site. Many commercial sequence analysis packages also provide search programs that use PROSITE data.

A resource for searching protein motifs is the BLOCKS E-mail server developed by Henikoff, Trends Biochem Sci. 18:267-268 (1993), the entirety of which is herein incorporated by reference; Henikoff and Henikoff, Nucleic Acid Research 19:6565-6572 (1991), the entirety of which is herein incorporated by reference; Henikoff and Henikoff, Proteins 17:49-61 (1993). BLOCKS searches a protein or nucleotide sequence against a database of protein motifs or “blocks.” Blocks are defined as short, ungapped multiple alignments that represent highly conserved protein patterns. The blocks themselves are derived from entries in PROSITE as well as other sources. Either a protein query or a nucleotide query can be submitted to the BLOCKS server; if a nucleotide sequence is submitted, the sequence is translated in all six reading frames and motifs are sought for these conceptual translations. Once the search is completed, the server will return a ranked list of significant matches, along with an alignment of the query sequence to the matched BLOCKS entries.

Conserved protein domains can be represented by two-dimensional matrices, which measure either the frequency or probability of the occurrences of each amino acid residue and deletions or insertions in each position of the domain. This type of model, when used to search against protein databases, is sensitive and usually yields more accurate results than simple motif searches. Two popular implementations of this approach are profile searches such as GCG program ProfileSearch and Hidden Markov Models (HMMs) (Krough et al., J. Mol. Biol. 235:1501-1531, (1994); Eddy, Current Opinion in Structural Biology 6:361-365, (1996), both of which are herein incorporated by reference in their entirety). In both cases, a large number of common protein domains have been converted into profiles, as present in the PROSITE library, or HHM models, as in the Pfam protein domain library (Sonnhammer et al., Proteins 28:405-420 (1997), the entirety of which is herein incorporated by reference). Pfam contains more than 500 HMM models for enzymes, transcription factors, signal transduction molecules and structural proteins. Protein databases can be queried with these profiles or HMM models, which will identify proteins containing the domain of interest. For example, HMMSW or HMMFS, two programs in a public domain package called HMMER (Sonnhammer et al., Proteins 28:405-420 (1997)) can be used.

PROSITE and BLOCKS represent collected families of protein motifs. Thus, searching these databases entails submitting a single sequence to determine whether or not that sequence is similar to the members of an established family. Programs working in the opposite direction compare a collection of sequences with individual entries in the protein databases. An example of such a program is the Motif Search Tool, or MoST (Tatusov et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:12091-12095 (1994), the entirety of which is herein incorporated by reference). On the basis of an aligned set of input sequences, a weight matrix is calculated by using one of four methods (selected by the user). A weight matrix is simply a representation, position by position of how likely a particular amino acid will appear. The calculated weight matrix is then used to search the databases. To increase sensitivity, newly found sequences are added to the original data set, the weight matrix is recalculated and the search is performed again. This procedure continues until no new sequences are found.

SUMMARY OF THE INVENTION

The present invention provides a substantially purified nucleic acid molecule that encodes a maize or soybean enzyme or fragment thereof, wherein said maize or soybean enzyme is selected from the group consisting of: (a) methionine adenosyltransferase, (b) S-adenosyl-methionine decarboxylase, (c) aspartate kinase, (d) aspartate-semialdehyde dehydrogenase, (e) cystathionine gamma-synthase, (f) cystathionine beta-lysase, and (g) 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase.

The present invention also provides a substantially purified nucleic acid molecule that encodes a plant methionine pathway enzyme or fragment thereof, wherein the nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine γ-lyase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof.

A substantially purified maize or soybean enzyme or fragment thereof, wherein said maize or soybean enzyme is selected from the group consisting of (a) methionine adenosyltransferase or fragment thereof; (b) S-adenosyl-methionine decarboxylase or fragment thereof; (c) aspartate kinase or fragment thereof; (d) aspartate-semialdehyde dehydrogenase or fragment thereof; (e) cystathionine gamma-synthase or fragment thereof; (f) cystathionine beta-lysase or fragment thereof; and (e) 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase or fragment thereof.

The present invention also provides a substantially purified maize or soybean methionine pathway protein or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 3204.

The present invention also provides a substantially purified maize or soybean methionine adenosyltransferase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO: 1635 through SEQ ID NO: 2479.

The present invention also provides a substantially purified maize or soybean methionine adenosyltransferase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO: 1635 through SEQ ID NO: 2479.

The present invention also provides a substantially purified maize or soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623.

The present invention also provides a substantially purified maize or soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623.

The present invention also provides a substantially purified maize or soybean aspartate kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO: 2648.

The present invention also provides a substantially purified maize or soybean aspartate kinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO: 2648.

The present invention also provides a substantially purified maize or soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO: 2654.

The present invention also provides a substantially purified maize or soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO: 2654.

The present invention also provides a substantially purified maize or soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO: 2660.

The present invention also provides a substantially purified maize or soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO: 2660.

The present invention also provides a substantially purified maize or soybean cystathionine β-lyase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2661 through SEQ ID NO: 2665.

The present invention also provides a substantially purified maize or soybean cystathionine β-lyase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2661 through SEQ ID NO: 2665.

The present invention also provides a substantially purified maize or soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 964 through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992.

The present invention also provides a substantially purified maize or soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 964 through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992.

The present invention also provides a substantially purified maize or adenosylhomocysteinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199.

The present invention also provides a substantially purified maize or adenosylhomocysteinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199.

The present invention also provides a substantially purified maize or cystathionine β-synthase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1631 through SEQ ID NO: 1632.

The present invention also provides a substantially purified maize or cystathionine β-synthase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1631 through SEQ ID NO: 1632.

The present invention also provides a substantially purified maize or cystathionine γ-lyase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204.

The present invention also provides a substantially purified maize or cystathionine γ-lyase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204.

The present invention also provides a substantially purified maize or O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 3200 through SEQ ID NO: 3202.

The present invention also provides a substantially purified maize or O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 3200 through SEQ ID NO: 3202.

The present invention also provides a purified antibody or fragment thereof which is capable of specifically binding to a specific maize or soybean enzyme or fragment thereof, wherein said maize or soybean enzyme or fragment thereof is encoded by a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of consisting of SEQ ID NO: 1 through SEQ ID NO: 3204.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean methionine adenosyltransferase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ. ID NO: 1635 through SEQ ID NO: 2479 or a substantially purified maize or soybean methionine adenosyltransferase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO: 1635 through SEQ ID NO: 2479.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623 or a substantially purified maize or soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean aspartate kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO: 2648 or a substantially purified maize or soybean aspartate kinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO: 2648.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO: 2654 or a substantially purified maize or soybean enzyme aspartate-semialdehyde dehydrogenase or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO: 2654.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO: 2660 or a substantially purified maize or soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO: 2660.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean cystathionine β-lyase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2661 through SEQ ID NO: 2665 or a substantially purified maize or soybean cystathionine β-lyase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2661 through SEQ ID NO: 2665.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 964 through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992 or a substantially purified maize or soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 964 through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean adenosylhomocyteinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199 or a substantially purified maize or soybean adenosylhomocyteinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean cystathionine β-synthase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1631 through SEQ ID NO: 1632 or a substantially purified maize or soybean cystathionine β-synthase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1631 through SEQ ID NO: 1632.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean cystathionine γ-lyase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204 or a substantially purified maize or soybean cystathionine γ-lyase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean O-acetylhomoserine enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 3200 through SEQ ID NO: 3202 or a substantially purified maize or soybean O-acetylhomoserine enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 3200 through SEQ ID NO: 3202.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; (B) a structural nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of (a) a nucleic acid sequence which encodes for methionine adenosyltransferase or fragment thereof, (b) a nucleic acid sequence which encodes for S-adenosyl-methionine decarboxylase or fragment thereof, (c) a nucleic acid sequence which encodes for aspartate kinase or fragment thereof; (d) a nucleic acid sequence which encodes for aspartate-semialdehyde dehydrogenase or fragment thereof; (e) a nucleic acid sequence which encodes for cystathionine gamma-synthase or a fragment thereof; (f) a nucleic acid sequence which encodes for cystathionine beta-lysase or a fragment thereof; (g) a nucleic acid sequence which encodes for 5-methyltetrahydropteroyl-triglutamate-homocysteine-S-methyltransferase or a fragment thereof; and (h) a nucleic acid sequence which is complementary to any of the nucleic acid sequences of (a) through (g); and (C) a 3′ non-translated sequence that functions in said plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of said mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a structural nucleic acid molecule, wherein the structural nucleic acid molecule encodes a plant methionine pathway enzyme or fragment thereof, the structural nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a structural nucleic acid molecule, wherein the structural nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or soybean aspartate kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine γ-lyase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a transcribed nucleic acid molecule with a transcribed strand and a non-transcribed strand, wherein the transcribed strand is complementary to a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in plant cells to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to: (B) a transcribed nucleic acid molecule with a transcribed strand and a non-transcribed strand, wherein a transcribed mRNA of the transcribed strand is complementary to an endogenous mRNA molecule having a nucleic acid sequence selected from the group consisting of an endogenous mRNA molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean aspartate kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean cystathionine γ-lyase enzyme or fragment thereof and an endogenous mRNA molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a method for determining a level or pattern in a plant cell of an enzyme in a plant metabolic pathway comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, said marker nucleic acid molecule selected from the group of marker nucleic acid molecules which specifically hybridize to a nucleic acid molecule having the nucleic acid sequence of SEQ ID NO: 1 through SEQ ID NO: 3204 or compliments thereof, with a complementary nucleic acid molecule obtained from said plant cell or plant tissue, wherein nucleic acid hybridization between said marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said plant cell or plant tissue permits the detection of an mRNA for said enzyme; (B) permitting hybridization between said marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said plant cell or plant tissue; and (C) detecting the level or pattern of said complementary nucleic acid, wherein the detection of said complementary nucleic acid is predictive of the level or pattern of said enzyme in said plant metabolic pathway.

The present invention also provides a method for determining a level or pattern of a plant methionine pathway enzyme in a plant cell or plant tissue comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complement thereof or fragment of either, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of the plant methionine pathway enzyme; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the plant methionine pathway enzyme.

The present invention also provides a method for determining a level or pattern of a plant methionine pathway enzyme in a plant cell or plant tissue comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean aspartate kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cytsathionine γ-lyase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complement thereof or fragment of either, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of the plant methionine pathway enzyme; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the plant methionine pathway enzyme.

The present invention also provides a method for determining a level or pattern of a plant methionine pathway enzyme in a plant cell or plant tissue under evaluation which comprises assaying the concentration of a molecule, whose concentration is dependent upon the expression of a gene, the gene specifically hybridizes to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof, in comparison to the concentration of that molecule present in a reference plant cell or a reference plant tissue with a known level or pattern of the plant methionine pathway enzyme, wherein the assayed concentration of the molecule is compared to the assayed concentration of the molecule in the reference plant cell or reference plant tissue with the known level or pattern of the plant methionine pathway enzyme.

The present invention also provides a method for determining a level or pattern of a plant methionine pathway enzyme in a plant cell or plant tissue under evaluation which comprises assaying the concentration of a molecule, whose concentration is dependent upon the expression of a gene, the gene specifically hybridizes to a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocyteinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine γ-lyase enzyme or complement thereof and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complement thereof, in comparison to the concentration of that molecule present in a reference plant cell or a reference plant tissue with a known level or pattern of the plant methionine pathway enzyme, wherein the assayed concentration of the molecule is compared to the assayed concentration of the molecule in the reference plant cell or the reference plant tissue with the known level or pattern of the plant methionine pathway enzyme.

A method of determining a mutation in a plant whose presence is predictive of a mutation affecting a level or pattern of a protein comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid, said marker nucleic acid selected from the group of marker nucleic acid molecules which specifically hybridize to a nucleic acid molecule having a nucleic acid sequence selected from the group of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof and a complementary nucleic acid molecule obtained from said plant, wherein nucleic acid hybridization between said marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting said level or pattern of said plant methionine pathway enzyme in said plant; (B) permitting hybridization between said marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said plant; and (C) detecting the presence of said polymorphism, wherein the detection of said polymorphism is predictive of said mutation.

The present invention also provides a method for determining a mutation in a plant whose presence is predictive of a mutation affecting the level or pattern of a plant methionine pathway enzyme comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that is linked to a gene, the gene specifically hybridizes to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant methionine pathway enzyme in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

The present invention also provides a method for determining a mutation in a plant whose presence is predictive of a mutation affecting the level or pattern of a plant methionine pathway enzyme comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that is linked to a gene, the gene specifically hybridizes to a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocyteinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine γ-lyase enzyme or complement thereof and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complement thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant methionine pathway enzyme in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

A method of producing a plant containing an overexpressed protein comprising: (A) transforming said plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein said promoter region is linked to a structural region, wherein said structural region has a nucleic acid sequence selected from group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 wherein said structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein said functional nucleic acid molecule results in overexpression of the protein; and (B) growing said transformed plant.

The present invention also provides a method of producing a plant containing an overexpressed plant methionine enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof; wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the plant methionine pathway enzyme; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing an overexpressed plant methionine pathway enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine γ-lyase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the plant methionine pathway enzyme protein; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing reduced levels of a plant methionine pathway enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204; wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in co-suppression of the plant methionine pathway enzyme protein; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing reduced levels of a plant methionine pathway enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine γ-lyase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in co-suppression of the plant methionine pathway enzyme; and (B) growing the transformed plant.

The present invention also provides a method for reducing expression of a plant methionine pathway enzyme in a plant comprising: (A) transforming the plant with a nucleic acid molecule, the nucleic acid molecule having an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule, wherein the exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein the transcribed strand is complementary to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof or fragments of either and the transcribed strand is complementary to an endogenous mRNA molecule; and wherein the transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and (B) growing the transformed plant.

The present invention also provides a method for reducing expression of a plant methionine pathway enzyme in a plant comprising: (A) transforming the plant with a nucleic acid molecule, the nucleic acid molecule having an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule, wherein the exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein a transcribed mRNA of the transcribed strand is complementary to a nucleic acid molecule selected from the group consisting of an endogenous mRNA molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean aspartate kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean cystathionine γ-lyase enzyme or fragment thereof and an endogenous mRNA molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; and wherein the transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and (B) growing the transformed plant.

The present invention also provides a method of determining an association between a polymorphism and a plant trait comprising: (A) hybridizing a nucleic acid molecule specific for the polymorphism to genetic material of a plant, wherein the nucleic acid molecule has a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof or fragment thereof; and (B) calculating the degree of association between the polymorphism and the plant trait.

The present invention also provides a method of determining an association between a polymorphism and a plant trait comprising: (A) hybridizing a nucleic acid molecule specific for the polymorphism to genetic material of a plant, wherein the nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean aspartate kinase enzyme complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or complement thereof or fragment of either; a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cytsathionine γ-lyase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complement thereof or fragment of either and (B) calculating the degree of association between the polymorphism and the plant trait.

The present invention also provides a method of isolating a nucleic acid that encodes a plant methionine pathway enzyme or fragment thereof comprising: (A) incubating under conditions permitting nucleic acid hybridization, a first nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof or fragment of either with a complementary second nucleic acid molecule obtained from a plant cell or plant tissue; (B) permitting hybridization between the first nucleic acid molecule and the second nucleic acid molecule obtained from the plant cell or plant tissue; and (C) isolating the second nucleic acid molecule.

The present invention also provides a method of isolating a nucleic acid molecule that encodes a plant methionine pathway enzyme or fragment thereof comprising: (A) incubating under conditions permitting nucleic acid hybridization, a first nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean aspartate kinase enzyme complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cytsathionine γ-lyase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complement thereof or fragment of either, with a complementary second nucleic acid molecule obtained from a plant cell or plant tissue; (B) permitting hybridization between the plant methionine pathway nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) isolating the second nucleic acid molecule.

DETAILED DESCRIPTION OF THE INVENTION Definitions and Agents of the Present Invention Definitions

As used herein, a methionine pathway enzyme is any enzyme that is associated with the synthesis or degradation of methionine.

As used herein, a methionine synthesis enzyme is any enzyme that is associated with the synthesis of methionine.

As used herein, a methionine degradation enzyme is any enzyme that is associated with the degradation of methionine.

As used herein, methionine adenosyltransferase is any enzyme that catalyzes the conversion of methionine to S-adenosylmethionine.

As used herein, S-adenosylmethionine decarboxylase is any enzyme that catalyzes the reaction that converts S-adenosylmethionine to (5-deoxy-5-adenosyl)(3-aminopropyl)methylsulfonuim salt.

As used herein, aspartate kinase is any enzyme that catalyzes the conversion of aspartate to β-aspartyl phosphate.

As used herein, aspartate semialdehyde dehydrogenase is any enzyme that catalyzes the conversion of β-aspartyl phosphate to aspartate-semialdehyde via an NADPH-dependent reaction.

As used herein, O-succinylhomoserine (thiol)-lyase refers to any enzyme that catalyzes the conversion of O-phosphohomoserine to and cysteine to cystathionine.

As used herein, cystathionine β-lyase is any enzyme that catalyzes the conversion of cystathionine to homocysteine, pyruvate and ammonia.

As used herein, 5-methyltetrhydropterolytriglutamate-homocysteine S-methyltransferase refers to any enzyme which catalyzes the conversion of homocysteine via methylation to methionine.

As used herein, adenosylhomocysteinase refers to any enzyme that catalyzes the ATP-dependent conversion of S-adenosylmethionine (AdoMet) to methylthioadenosine and L-homoserine.

As used herein, cystathionine β-synthase refers to any enzyme that catalyzes the conversion of homocysteine and serine to cystathionine.

As used herein, cystathionine γ-lyase refers to any enzyme that catalyzes the γ cleavage of cystationine.

As used herein, O-acetyhomoserine (thiol)-lyase refers to any enzyme that catalyzes the conversion of O-acetylhomoserine and sulfur to homocysteine.

Agents

(a) Nucleic Acid Molecules

Agents of the present invention include plant nucleic acid molecules and more specifically include maize and soybean nucleic acid molecules and more specifically include nucleic acid molecules of the maize genotypes B73 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), B73 x Mo17 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), DK604 (Dekalb Genetics, Dekalb, Ill. U.S.A.), H99 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), RX601 (Asgrow Seed Company, Des Moines, Iowa), Mo17 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), and soybean types Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa), C1944 (United States Department of Agriculture (USDA) Soybean Germplasm Collection, Urbana, Ill. U.S.A.), Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), FT108 (Monsoy, Brazil), Hartwig (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), BW211S Null (Tohoku University, Morioka, Japan), PI507354 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), Asgrow A4922 (Asgrow Seed Company, Des Moines, Iowa U.S.A.), PI227687 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), PI229358 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and Asgrow A3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.).

A subset of the nucleic acid molecules of the present invention includes nucleic acid molecules that are marker molecules. Another subset of the nucleic acid molecules of the present invention include nucleic acid molecules that encode a protein or fragment thereof. Another subset of the nucleic acid molecules of the present invention are EST molecules.

Fragment nucleic acid molecules may encode significant portion(s) of, or indeed most of, these nucleic acid molecules. Alternatively, the fragments may comprise smaller oligonucleotides (having from about 15 to about 250 nucleotide residues and more preferably, about 15 to about 30 nucleotide residues).

As used herein, an agent, be it a naturally occurring molecule or otherwise may be “substantially purified,” if desired, such that one or more molecules that is or may be present in a naturally occurring preparation containing that molecule will have been removed or will be present at a lower concentration than that at which it would normally be found.

The agents of the present invention will preferably be “biologically active” with respect to either a structural attribute, such as the capacity of a nucleic acid to hybridize to another nucleic acid molecule, or the ability of a protein to be bound by an antibody (or to compete with another molecule for such binding). Alternatively, such an attribute may be catalytic and thus involve the capacity of the agent to mediate a chemical reaction or response.

The agents of the present invention may also be recombinant. As used herein, the term recombinant means any agent (e.g. DNA, peptide etc.), that is, or results, however indirect, from human manipulation of a nucleic acid molecule.

It is understood that the agents of the present invention may be labeled with reagents that facilitate detection of the agent (e.g. fluorescent labels, Prober et al., Science 238:336-340 (1987); Albarella et al., EP 144914; chemical labels, Sheldon et al., U.S. Pat. No. 4,582,789; Albarella et al., U.S. Pat. No. 4,563,417; modified bases, Miyoshi et al., EP 119448, all of which are hereby incorporated by reference in their entirety).

It is further understood, that the present invention provides recombinant bacterial, mammalian, microbial, insect, fungal and plant cells and viral constructs comprising the agents of the present invention. (See, for example, Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants; Section (b) Fungal Constructs and Fungal Transformants; Section (c) Mammalian Constructs and Transformed Mammalian Cells; Section (d) Insect Constructs and Transformed Insect Cells; and Section (e) Bacterial Constructs and Transformed Bacterial Cells)

Nucleic acid molecules or fragments thereof of the present invention are capable of specifically hybridizing to other nucleic acid molecules under certain circumstances. As used herein, two nucleic acid molecules are said to be capable of specifically hybridizing to one another if the two molecules are capable of forming an anti-parallel, double-stranded nucleic acid structure. A nucleic acid molecule is said to be the “complement” of another nucleic acid molecule if they exhibit complete complementarity. As used herein, molecules are said to exhibit “complete complementarity” when every nucleotide of one of the molecules is complementary to a nucleotide of the other. Two molecules are said to be “minimally complementary” if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under at least conventional “low-stringency” conditions. Similarly, the molecules are said to be “complementary” if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under conventional “high-stringency” conditions. Conventional stringency conditions are described by Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd Ed., Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989) and by Haymes et al., Nucleic Acid Hybridization, A Practical Approach, IRL Press, Washington, D.C. (1985), the entirety of which is herein incorporated by reference. Departures from complete complementarity are therefore permissible, as long as such departures do not completely preclude the capacity of the molecules to form a double-stranded structure. Thus, in order for a nucleic acid molecule to serve as a primer or probe it need only be sufficiently complementary in sequence to be able to form a stable double-stranded structure under the particular solvent and salt concentrations employed.

Appropriate stringency conditions which promote DNA hybridization, for example, 6.0× sodium chloride/sodium citrate (SSC) at about 45° C., followed by a wash of 2.0×SSC at 50° C., are known to those skilled in the art or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. For example, the salt concentration in the wash step can be selected from a low stringency of about 2.0×SSC at 50° C. to a high stringency of about 0.2×SSC at 50° C. In addition, the temperature in the wash step can be increased from low stringency conditions at room temperature, about 22° C., to high stringency conditions at about 65° C. Both temperature and salt may be varied, or either the temperature or the salt concentration may be held constant while the other variable is changed.

In a preferred embodiment, a nucleic acid of the present invention will specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof under moderately stringent conditions, for example at about 2.0×SSC and about 65° C.

In a particularly preferred embodiment, a nucleic acid of the present invention will include those nucleic acid molecules that specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof under high stringency conditions such as 0.2×SSC and about 65° C.

In one aspect of the present invention, the nucleic acid molecules of the present invention have one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof. In another aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 90% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof. In a further aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 95% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof. In a more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 98% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof. In an even more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 99% sequence identity with one or more of the sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof.

In a further more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention exhibit 100% sequence identity with a nucleic acid molecule present within MONN01, SATMON001 through SATMON031, SATMON033, SATMON034, SATMON˜001, SATMONN01, SATMONN04 through SATMONN006, CMz029 through CMz031, CMz033, CMz035 through CMz037, CMz039 through CMz042, CMz044 through CMz045, CMz047 through CMz050, SOYMON001 through SOYMON038, Soy51 through Soy56, Soy58 through Soy62, Soy65 through Soy66, Soy 68 through Soy73 and Soy76 through Soy77, Lib9, Lib22 through Lib25, Lib35, Lib80 through Lib81, Lib 144, Lib146, Lib147, Lib190, Lib3032 through Lib3036 and Lib3099 (Monsanto Company, St. Louis, Mo. U.S.A.).

(i) Nucleic Acid Molecules Encoding Proteins or Fragments Thereof

Nucleic acid molecules of the present invention can comprise sequences that encode a methionine pathway protein or fragment thereof. Such proteins or fragments thereof include homologues of known proteins in other organisms.

In a preferred embodiment of the present invention, a maize or soybean protein homologue or fragment thereof of the present invention is a homologue of another plant protein. In another preferred embodiment of the present invention, a maize or soybean protein homologue or fragment thereof of the present invention is a homologue of a fungal protein. In another preferred embodiment of the present invention, a maize or soybean protein homologue of the present invention is a homologue of mammalian protein. In another preferred embodiment of the present invention, a maize or soybean protein homologue or fragment thereof of the present invention is a homologue of a bacterial protein. In another preferred embodiment of the present invention, a soybean protein homologue or fragment thereof of the present invention is a homologue of a maize protein. In another preferred embodiment of the present invention, a maize protein homologue or fragment thereof of the present invention is a homologue of a soybean protein.

In a preferred embodiment of the present invention, the nucleic molecule of the present invention encodes a maize or soybean homologue protein or fragment thereof where a maize or soybean homologue protein exhibits a BLAST probability score of greater than 1E-12, preferably a BLAST probability score of between about 1E-30 and about 1E-12, even more preferably a BLAST probability score of greater than 1E-30 with its homologue.

In another preferred embodiment of the present invention, the nucleic acid molecule encoding a maize or soybean protein homologue or fragment thereof or fragment thereof exhibits a % identity with its homologue of between about 25% and about 40%, more preferably of between about 40 and about 70%, even more preferably of between about 70% and about 90% and even more preferably between about 90% and 99%. In another preferred embodiment, of the present invention, a maize or soybean protein homologue or fragments thereof exhibits a % identity with its homologue of 100%.

In a preferred embodiment of the present invention, the nucleic molecule of the present invention encodes a maize or soybean homologue protein or fragment thereof where a maize or soybean homologue protein exhibits a BLAST score of greater than 120, preferably a BLAST score of between about 1450 and about 120, even more preferably a BLAST score of greater than 1450 with its homologue.

Nucleic acid molecules of the present invention also include non-maize, non-soybean homologues. Preferred non-homologues are selected from the group consisting of alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm and Phaseolus.

In a preferred embodiment, nucleic acid molecules having SEQ ID NO: 1 through SEQ ID NO: 3204 or complements and fragments of either can be utilized to obtain such homologues.

The degeneracy of the genetic code, which allows different nucleic acid sequences to code for the same protein or peptide, is known in the literature. (U.S. Pat. No. 4,757,006, the entirety of which is herein incorporated by reference).

In an aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding maize or soybean homologue or fragment thereof in SEQ ID NO: 1 through SEQ ID NO: 3204 due to the degeneracy in the genetic code in that they encode the same protein but differ in nucleic acid sequence.

In another further aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding maize or soybean homologue or fragment thereof in SEQ ID NO: 1 through SEQ ID NO: 3204 due to fact that the different nucleic acid sequence encodes a protein having one or more conservative amino acid residue. Examples of conservative substitutions are set forth in Table 1. It is understood that codons capable of coding for such conservative substitutions are known in the art.

TABLE 1 Original Residue Conservative Substitutions Ala Ser Arg Lys Asn Gln; His Asp Glu Cys Ser; Ala Gln Asn Glu Asp Gly Pro His Asn; Gln Ile Leu; Val Leu Ile; Val Lys Arg; Gln; Glu Met Leu; Ile Phe Met; Leu; Tyr Ser Thr Thr Ser Trp Tyr Tyr Trp; Phe Val Ile; Leu

In a further aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or soybean homologue or fragment thereof set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof due to the fact that one or more codons encoding an amino acid has been substituted for a codon that encodes a nonessential substitution of the amino acid originally encoded.

Agents of the present invention include nucleic acid molecules that encode a maize or soybean methionine pathway protein or fragment thereof and particularly substantially purified nucleic acid molecules selected from the group consisting of a nucleic acid molecule that encodes a maize or soybean methionine adenosyltransferase protein or fragment thereof, a nucleic acid molecule that encodes a maize or soybean S-adenosylmethionine decarboxylase protein or fragment thereof, a nucleic acid molecule that encodes a maize or soybean aspartate kinase protein or fragment thereof, a nucleic acid molecule that encode a maize or soybean aspartate-semialdehyde dehydrogenase protein or fragment thereof, a nucleic acid molecule that encodes a maize or soybean O-succinylhomoserine (thiol)-lyase protein or fragment thereof, a nucleic acid molecule that encodes a maize or soybean cystathionine β-lyase protein or fragment thereof, a nucleic acid molecule that encodes a maize or soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase protein or fragment thereof, a nucleic acid molecule that encodes a maize or soybean adenosylhomocysteine protein or fragment thereof, a nucleic acid molecule that encodes a maize or soybean cystathionine β-synthase protein or fragment thereof, a nucleic acid molecule that encodes a maize or soybean cystathionine γ-lyase protein or fragment thereof, and a nucleic acid molecule that encodes a maize or soybean O-acetylhomoserine (thiol)-lyase protein or fragment thereof.

Non-limiting examples of such nucleic acid molecules of the present invention are nucleic acid molecules comprising: SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof that encode for a methionine pathway protein or fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO: 1635 through SEQ ID NO: 2479 or fragment thereof that encode for a methionine adenosyltransferase protein or fragment thereof, SEQ ID NO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623 or fragment thereof that encode for a S-adenosylmethionine decarboxylase protein or fragment thereof, SEQ ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO: 2648 or fragment thereof that encode for a aspartate kinase protein or fragment thereof, SEQ ID NO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO: 2654 or fragment thereof that encode for a aspartate-semialdehyde dehydrogenase protein or fragment thereof, SEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO: 2660 or fragment thereof that encode for a O-succinylhomoserine (thiol)-lyase protein or fragment thereof, SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2655 through SEQ ID NO: 2660 or fragment thereof that encode for a cystathionine β-lyase protein or fragment thereof, SEQ ID NO: 964 through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992 or fragment thereof that encode for a 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase protein or fragment thereof, SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199 or fragment thereof that encode for an adenosylhomocysteinase protein or fragment thereof, SEQ ID NO: 1631 through SEQ ID NO: 1632 or fragment thereof that encode for a cystathionine β-synthase protein or fragment thereof, SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204 or fragment thereof that encode for a cystathionine γ-lyase protein or fragment thereof, and SEQ ID NO: 3200 through SEQ ID NO: 3202 or fragment thereof that encode for an O-acetylhomoserine (thiol)-lyase protein or fragment thereof.

A nucleic acid molecule of the present invention can also encode an homologue of a maize or soybean methionine adenosyltransferase or fragment thereof, a maize or soybean S-adenosylmethionine decarboxylase or fragment thereof, a maize or soybean aspartate kinase or fragment thereof, a maize or soybean aspartate-semialdehyde dehydrogenase or fragment thereof, a maize or soybean O-succinylhomoserine (thiol)-lyase or fragment thereof, a maize or soybean cystathionine β-lyase or fragment thereof, a maize or soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase or fragment thereof, a maize or soybean adenosylhomocysteinease or fragment thereof, a maize or soybean cystathionine β-synthase or fragment thereof, a maize or soybean cystathionine γ-lyase or fragment thereof or a maize or soybean O-acetylhomoserine (thiol)-lyase or fragment thereof. As used herein a homologue protein molecule or fragment thereof is a counterpart protein molecule or fragment thereof in a second species (e.g., maize methionine adenosyltransferase protein is a homologue of Arabidopsis' methionine adenosyltransferase protein).

(ii) Nucleic Acid Molecule Markers and Probes

One aspect of the present invention concerns markers that include nucleic acid molecules SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof or fragments of either that can act as markers. Genetic markers of the present invention include “dominant” or “codominant” markers. “Codominant markers” reveal the presence of two or more alleles (two per diploid individual) at a locus. “Dominant markers” reveal the presence of only a single allele per locus. The presence of the dominant marker phenotype (e.g., a band of DNA) is an indication that one allele is present in either the homozygous or heterozygous condition. The absence of the dominant marker phenotype (e.g. absence of a DNA band) is merely evidence that “some other” undefined allele is present. In the case of populations where individuals are predominantly homozygous and loci are predominately dimorphic, dominant and codominant markers can be equally valuable. As populations become more heterozygous and multi-allelic, codominant markers often become more informative of the genotype than dominant markers. Marker molecules can be, for example, capable of detecting polymorphisms such as single nucleotide polymorphisms (SNPs).

SNPs are single base changes in genomic DNA sequence. They occur at greater frequency and are spaced with a greater uniformly throughout a genome than other reported forms of polymorphism. The greater frequency and uniformity of SNPs means that there is greater probability that such a polymorphism will be found near or in a genetic locus of interest than would be the case for other polymorphisms. SNPs are located in protein-coding regions and noncoding regions of a genome. Some of these SNPs may result in defective or variant protein expression (e.g., as a results of mutations or defective splicing). Analysis (genotyping) of characterized SNPs can require only a plus/minus assay rather than a lengthy measurement, permitting easier automation.

SNPs can be characterized using any of a variety of methods. Such methods include the direct or indirect sequencing of the site, the use of restriction enzymes (Botstein et al., Am. J. Hum. Genet. 32:314-331 (1980), the entirety of which is herein incorporated reference; Konieczny and Ausubel, Plant J. 4:403-410 (1993), the entirety of which is herein incorporated by reference), enzymatic and chemical mismatch assays (Myers et al., Nature 313:495-498 (1985), the entirety of which is herein incorporated by reference), allele-specific PCR (Newton et al., Nucl. Acids Res. 17:2503-2516 (1989), the entirety of which is herein incorporated by reference; Wu et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:2757-2760 (1989), the entirety of which is herein incorporated by reference), ligase chain reaction (Barany, Proc. Natl. Acad. Sci. (U.S.A.) 88:189-193 (1991), the entirety of which is herein incorporated by reference), single-strand conformation polymorphism analysis (Labrune et al., Am. J. Hum. Genet. 48: 1115-1120 (1991), the entirety of which is herein incorporated by reference), primer-directed nucleotide incorporation assays (Kuppuswami et al., Proc. Natl. Acad. Sci. USA 88:1143-1147 (1991), the entirety of which is herein incorporated by reference), dideoxy fingerprinting (Sarkar et al., Genomics 13:441-443 (1992), the entirety of which is herein incorporated by reference), solid-phase ELISA-based oligonucleotide ligation assays (Nikiforov et al., Nucl. Acids Res. 22:4167-4175 (1994), the entirety of which is herein incorporated by reference), oligonucleotide fluorescence-quenching assays (Livak et al., PCR Methods Appl. 4:357-362 (1995), the entirety of which is herein incorporated by reference), 5′-nuclease allele-specific hybridization TaqMan assay (Livak et al., Nature Genet. 9:341-342 (1995), the entirety of which is herein incorporated by reference), template-directed dye-terminator incorporation (TDI) assay (Chen and Kwok, Nucl. Acids Res. 25:347-353 (1997), the entirety of which is herein incorporated by reference), allele-specific molecular beacon assay (Tyagi et al., Nature Biotech. 16: 49-53 (1998), the entirety of which is herein incorporated by reference), PinPoint assay (Haff and Smimov, Genome Res. 7: 378-388 (1997), the entirety of which is herein incorporated by reference) and dCAPS analysis (Neff et al., Plant J. 14:387-392 (1998), the entirety of which is herein incorporated by reference).

Additional markers, such as AFLP markers, RFLP markers and RAPD markers, can be utilized (Walton, Seed World 22-29 (July, 1993), the entirety of which is herein incorporated by reference; Burow and Blake, Molecular Dissection of Complex Traits, 13-29, Paterson (ed.), CRC Press, New York (1988), the entirety of which is herein incorporated by reference). DNA markers can be developed from nucleic acid molecules using restriction endonucleases, the PCR and/or DNA sequence information. RFLP markers result from single base changes or insertions/deletions. These codominant markers are highly abundant in plant genomes, have a medium level of polymorphism and are developed by a combination of restriction endonuclease digestion and Southern blotting hybridization. CAPS are similarly developed from restriction nuclease digestion but only of specific PCR products. These markers are also codominant, have a medium level of polymorphism and are highly abundant in the genome. The CAPS result from single base changes and insertions/deletions.

Another marker type, RAPDs, are developed from DNA amplification with random primers and result from single base changes and insertions/deletions in plant genomes. They are dominant markers with a medium level of polymorphisms and are highly abundant. AFLP markers require using the PCR on a subset of restriction fragments from extended adapter primers. These markers are both dominant and codominant are highly abundant in genomes and exhibit a medium level of polymorphism.

SSRs require DNA sequence information. These codominant markers result from repeat length changes, are highly polymorphic and do not exhibit as high a degree of abundance in the genome as CAPS, AFLPs and RAPDs SNPs also require DNA sequence information. These codominant markers result from single base substitutions. They are highly abundant and exhibit a medium of polymorphism (Rafalski et al., In: Nonmammalian Genomic Analysis, Birren and Lai (ed.), Academic Press, San Diego, Calif., pp. 75-134 (1996), the entirety of which is herein incorporated by reference). It is understood that a nucleic acid molecule of the present invention may be used as a marker.

A PCR probe is a nucleic acid molecule capable of initiating a polymerase activity while in a double-stranded structure to with another nucleic acid. Various methods for determining the structure of PCR probes and PCR techniques exist in the art. Computer generated searches using programs such as Primer3 (on the Worldwide web at genome.wi.mit.edu/cgi-bin/primer/primer3.cgi), STSPipeline (on the Worldwide web at genome.wi.mit.edu/cgi-bin/www-STS_Pipeline), or GeneUp (Pesole et al., Bio Techniques 25:112-123 (1998) the entirety of which is herein incorporated by reference), for example, can be used to identify potential PCR primers.

It is understood that a fragment of one or more of the nucleic acid molecules of the present invention may be a probe and specifically a PCR probe.

(b) Protein and Peptide Molecules

A class of agents comprises one or more of the protein or fragments thereof or peptide molecules encoded by SEQ ID NO: 1 through SEQ ID NO: 3204 or one or more of the protein or fragment thereof and peptide molecules encoded by other nucleic acid agents of the present invention. As used herein, the term “protein molecule” or “peptide molecule” includes any molecule that comprises five or more amino acids. It is well known in the art that proteins may undergo modification, including post-translational modifications, such as, but not limited to, disulfide bond formation, glycosylation, phosphorylation, or oligomerization. Thus, as used herein, the term “protein molecule” or “peptide molecule” includes any protein molecule that is modified by any biological or non-biological process. The terms “amino acid” and “amino acids” refer to all naturally occurring L-amino acids. This definition is meant to include norleucine, ornithine, homocysteine and homoserine.

Non-limiting examples of the protein or fragment thereof of the present invention include a maize or soybean methionine pathway protein or fragment thereof; a maize or soybean methionine adenosyltransferase or fragment thereof, a maize or soybean S-adenosylmethionine decarboxylase or fragment thereof, a maize or soybean aspartate kinase or fragment thereof, a maize or soybean aspartate-semialdehyde dehydrogenase or fragment thereof, a maize or soybean O-succinylhomoserine (thiol)-lyase or fragment thereof, a maize or soybean cystathionine β-lyase or fragment thereof, a maize or soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase or fragment thereof, a maize or soybean adenosylhomocysteinase or fragment thereof, a maize or soybean cystathionine β-synthase or fragment thereof, a maize or soybean cystathionine γ-lyase or fragment thereof or a maize or soybean O-acetylhomoserine (thiol)-lyase or fragment thereof.

Non-limiting examples of the protein or fragment molecules of the present invention are an methionine pathway protein or fragment thereof encoded by: SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof that encode for a methionine pathway protein or fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO: 1635 through SEQ ID NO: 2479 or fragment thereof that encode for a methionine adenosyltransferase protein or fragment thereof, SEQ ID NO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623 or fragment thereof that encode for a S-adenosylmethionine decarboxylase protein or fragment thereof, SEQ ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO: 2648 or fragment thereof that encode for a aspartate kinase protein or fragment thereof, SEQ ID NO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO: 2654 or fragment thereof that encode for a aspartate-semialdehyde dehydrogenase protein or fragment thereof, SEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO: 2660 or fragment thereof that encode for a O-succinylhomoserine (thiol)-lyase protein or fragment thereof, SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2655 through SEQ ID NO: 2660 or fragment thereof that encode for a cystathionine β-lyase or fragment thereof, SEQ ID NO: 964 through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992 or fragment thereof that encode for a 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase protein or fragment thereof, SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199 or fragment thereof that encode for an adenosylhomocysteinase protein or fragment thereof, SEQ ID NO: 1631 through SEQ ID NO: 1632 or fragment thereof that encode for a cystathionine β-synthase protein or fragment thereof, SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204 or fragment thereof that encode for a cystathionine γ-lyase protein or fragment thereof, and SEQ ID NO: 3200 through SEQ ID NO: 3202 or fragment thereof that encode for an O-acetylhomoserine (thiol)-lyase protein or fragment thereof.

One or more of the protein or fragment of peptide molecules may be produced via chemical synthesis, or more preferably, by expressing in a suitable bacterial or eucaryotic host. Suitable methods for expression are described by Sambrook et al., (In: Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989)), or similar texts. For example, the protein may be expressed in, for example, Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants; Section (b) Fungal Constructs and Fungal Transformants; Section (c) Mammalian Constructs and Transformed Mammalian Cells; Section (d) Insect Constructs and Transformed Insect Cells; and Section (e) Bacterial Constructs and Transformed Bacterial Cells.

A “protein fragment” is a peptide or polypeptide molecule whose amino acid sequence comprises a subset of the amino acid sequence of that protein. A protein or fragment thereof that comprises one or more additional peptide regions not derived from that protein is a “fusion” protein. Such molecules may be derivatized to contain carbohydrate or other moieties (such as keyhole limpet hemocyanin, etc.). Fusion protein or peptide molecules of the present invention are preferably produced via recombinant means.

Another class of agents comprise protein or peptide molecules or fragments or fusions thereof encoded by SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof in which conservative, non-essential or non-relevant amino acid residues have been added, replaced or deleted. Computerized means for designing modifications in protein structure are known in the art (Dahiyat and Mayo, Science 278:82-87 (1997), the entirety of which is herein incorporated by reference).

The protein molecules of the present invention include plant homologue proteins. An example of such a homologue is a homologue protein of a non-maize or non soybean plant species, that include but not limited to alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm, Phaseolus etc. Particularly preferred non-maize or non-soybean for use for the isolation of homologs would include, Arabidopsis, barley, cotton, oat, oilseed rape, rice, canola, ornamentals, sugarcane, sugarbeet, tomato, potato, wheat and turf grasses. Such a homologue can be obtained by any of a variety of methods. Most preferably, as indicated above, one or more of the disclosed sequences (SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof) will be used to define a pair of primers that may be used to isolate the homologue-encoding nucleic acid molecules from any desired species. Such molecules can be expressed to yield homologues by recombinant means.

(c) Antibodies

One aspect of the present invention concerns antibodies, single-chain antigen binding molecules, or other proteins that specifically bind to one or more of the protein or peptide molecules of the present invention and their homologues, fusions or fragments. Such antibodies may be used to quantitatively or qualitatively detect the protein or peptide molecules of the present invention. As used herein, an antibody or peptide is said to “specifically bind” to a protein or peptide molecule of the present invention if such binding is not competitively inhibited by the presence of non-related molecules.

Nucleic acid molecules that encode all or part of the protein of the present invention can be expressed, via recombinant means, to yield protein or peptides that can in turn be used to elicit antibodies that are capable of binding the expressed protein or peptide. Such antibodies may be used in immunoassays for that protein. Such protein-encoding molecules, or their fragments may be a “fusion” molecule (i.e., a part of a larger nucleic acid molecule) such that, upon expression, a fusion protein is produced. It is understood that any of the nucleic acid molecules of the present invention may be expressed, via recombinant means, to yield proteins or peptides encoded by these nucleic acid molecules.

The antibodies that specifically bind proteins and protein fragments of the present invention may be polyclonal or monoclonal and may comprise intact immunoglobulins, or antigen binding portions of immunoglobulins fragments (such as (F(ab′), F(ab′)₂), or single-chain immunoglobulins producible, for example, via recombinant means. It is understood that practitioners are familiar with the standard resource materials which describe specific conditions and procedures for the construction, manipulation and isolation of antibodies (see, for example, Harlow and Lane, In: Antibodies: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1988), the entirety of which is herein incorporated by reference).

Murine monoclonal antibodies are particularly preferred. BALB/c mice are preferred for this purpose, however, equivalent strains may also be used. The animals are preferably immunized with approximately 25 μg of purified protein (or fragment thereof) that has been emulsified in a suitable adjuvant (such as TiterMax adjuvant (Vaxcel, Norcross, Ga.)). Immunization is preferably conducted at two intramuscular sites, one intraperitoneal site and one subcutaneous site at the base of the tail. An additional i.v. injection of approximately 25 μg of antigen is preferably given in normal saline three weeks later. After approximately 11 days following the second injection, the mice may be bled and the blood screened for the presence of anti-protein or peptide antibodies. Preferably, a direct binding Enzyme-Linked Immunoassay (ELISA) is employed for this purpose.

More preferably, the mouse having the highest antibody titer is given a third i.v. injection of approximately 25 μg of the same protein or fragment. The splenic leukocytes from this animal may be recovered 3 days later and then permitted to fuse, most preferably, using polyethylene glycol, with cells of a suitable myeloma cell line (such as, for example, the P3X63Ag8.653 myeloma cell line). Hybridoma cells are selected by culturing the cells under “HAT” (hypoxanthine-aminopterin-thymine) selection for about one week. The resulting clones may then be screened for their capacity to produce monoclonal antibodies (“mAbs”), preferably by direct ELISA.

In one embodiment, anti-protein or peptide monoclonal antibodies are isolated using a fusion of a protein or peptide of the present invention, or conjugate of a protein or peptide of the present invention, as immunogens. Thus, for example, a group of mice can be immunized using a fusion protein emulsified in Freund's complete adjuvant (e.g. approximately 50 μg of antigen per immunization). At three week intervals, an identical amount of antigen is emulsified in Freund's incomplete adjuvant and used to immunize the animals. Ten days following the third immunization, serum samples are taken and evaluated for the presence of antibody. If antibody titers are too low, a fourth booster can be employed. Polysera capable of binding the protein or peptide can also be obtained using this method.

In a preferred procedure for obtaining monoclonal antibodies, the spleens of the above-described immunized mice are removed, disrupted and immune splenocytes are isolated over a ficoll gradient. The isolated splenocytes are fused, using polyethylene glycol with BALB/c-derived HGPRT (hypoxanthine guanine phosphoribosyl transferase) deficient P3x63xAg8.653 plasmacytoma cells. The fused cells are plated into 96 well microtiter plates and screened for hybridoma fusion cells by their capacity to grow in culture medium supplemented with hypothanthine, aminopterin and thymidine for approximately 2-3 weeks.

Hybridoma cells that arise from such incubation are preferably screened for their capacity to produce an immunoglobulin that binds to a protein of interest. An indirect ELISA may be used for this purpose. In brief, the supernatants of hybridomas are incubated in microtiter wells that contain immobilized protein. After washing, the titer of bound immunoglobulin can be determined using, for example, a goat anti-mouse antibody conjugated to horseradish peroxidase. After additional washing, the amount of immobilized enzyme is determined (for example through the use of a chromogenic substrate). Such screening is performed as quickly as possible after the identification of the hybridoma in order to ensure that a desired clone is not overgrown by non-secreting neighbor cells. Desirably, the fusion plates are screened several times since the rates of hybridoma growth vary. In a preferred sub-embodiment, a different antigenic form may be used to screen the hybridoma. Thus, for example, the splenocytes may be immunized with one immunogen, but the resulting hybridomas can be screened using a different immunogen. It is understood that any of the protein or peptide molecules of the present invention may be used to raise antibodies.

As discussed below, such antibody molecules or their fragments may be used for diagnostic purposes. Where the antibodies are intended for diagnostic purposes, it may be desirable to derivatize them, for example with a ligand group (such as biotin) or a detectable marker group (such as a fluorescent group, a radioisotope or an enzyme).

The ability to produce antibodies that bind the protein or peptide molecules of the present invention permits the identification of mimetic compounds of those molecules. A “mimetic compound” is a compound that is not that compound, or a fragment of that compound, but which nonetheless exhibits an ability to specifically bind to antibodies directed against that compound.

It is understood that any of the agents of the present invention can be substantially purified and/or be biologically active and/or recombinant.

Uses of the Agents of the Invention

Nucleic acid molecules and fragments thereof of the present invention may be employed to obtain other nucleic acid molecules from the same species (e.g., ESTs or fragment thereof from maize may be utilized to obtain other nucleic acid molecules from maize). Such nucleic acid molecules include the nucleic acid molecules that encode the complete coding sequence of a protein and promoters and flanking sequences of such molecules. In addition, such nucleic acid molecules include nucleic acid molecules that encode for other isozymes or gene family members. Such molecules can be readily obtained by using the above-described nucleic acid molecules or fragments thereof to screen cDNA or genomic libraries obtained from maize or soybean. Methods for forming such libraries are well known in the art.

Nucleic acid molecules and fragments thereof of the present invention may also be employed to obtain nucleic acid homologues. Such homologues include the nucleic acid molecule of other plants or other organisms (e.g., alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm, Phaseolus, etc.) including the nucleic acid molecules that encode, in whole or in part, protein homologues of other plant species or other organisms, sequences of genetic elements such as promoters and transcriptional regulatory elements. Such molecules can be readily obtained by using the above-described nucleic acid molecules or fragments thereof to screen cDNA or genomic libraries obtained from such plant species. Methods for forming such libraries are well known in the art. Such homologue molecules may differ in their nucleotide sequences from those found in one or more of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof because complete complementarity is not needed for stable hybridization. The nucleic acid molecules of the present invention therefore also include molecules that, although capable of specifically hybridizing with the nucleic acid molecules may lack “complete complementarity.”

Any of a variety of methods may be used to obtain one or more of the above-described nucleic acid molecules (Zamechik et al., Proc. Natl. Acad. Sci. (U.S.A.) 83:4143-4146 (1986), the entirety of which is herein incorporated by reference; Goodchild et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:5507-5511 (1988), the entirety of which is herein incorporated by reference; Wickstrom et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:1028-1032 (1988), the entirety of which is herein incorporated by reference; Holt et al., Molec. Cell. Biol. 8:963-973 (1988), the entirety of which is herein incorporated by reference; Gerwirtz et al., Science 242:1303-1306 (1988), the entirety of which is herein incorporated by reference; Anfossi et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:3379-3383 (1989), the entirety of which is herein incorporated by reference; Becker et al., EMBO J. 8:3685-3691 (1989); the entirety of which is herein incorporated by reference). Automated nucleic acid synthesizers may be employed for this purpose. In lieu of such synthesis, the disclosed nucleic acid molecules may be used to define a pair of primers that can be used with the polymerase chain reaction (Mullis et al., Cold Spring Harbor Symp. Quant. Biol. 51:263-273 (1986); Erlich et al., European Patent 50,424; European Patent 84,796; European Patent 258,017; European Patent 237,362; Mullis, European Patent 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194, all of which are herein incorporated by reference in their entirety) to amplify and obtain any desired nucleic acid molecule or fragment.

Promoter sequence(s) and other genetic elements, including but not limited to transcriptional regulatory flanking sequences, associated with one or more of the disclosed nucleic acid sequences can also be obtained using the disclosed nucleic acid sequence provided herein. In one embodiment, such sequences are obtained by incubating EST nucleic acid molecules or preferably fragments thereof with members of genomic libraries (e.g. maize and soybean) and recovering clones that hybridize to the EST nucleic acid molecule or fragment thereof. In a second embodiment, methods of “chromosome walking,” or inverse PCR may be used to obtain such sequences (Frohman et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:8998-9002 (1988); Ohara et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:5673-5677 (1989); Pang et al., Biotechniques 22:1046-1048 (1977); Huang et al., Methods Mol. Biol. 69:89-96 (1997); Huang et al., Method Mol. Biol. 67:287-294 (1997); Benkel et al., Genet. Anal. 13:123-127 (1996); Hartl et al., Methods Mol. Biol. 58:293-301 (1996), all of which are herein incorporated by reference in their entirety).

The nucleic acid molecules of the present invention may be used to isolate promoters of cell enhanced, cell specific, tissue enhanced, tissue specific, developmentally or environmentally regulated expression profiles. Isolation and functional analysis of the 5′ flanking promoter sequences of these genes from genomic libraries, for example, using genomic screening methods and PCR techniques would result in the isolation of useful promoters and transcriptional regulatory elements. These methods are known to those of skill in the art and have been described (See, for example, Birren et al., Genome Analysis: Analyzing DNA, 1, (1997), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., the entirety of which is herein incorporated by reference). Promoters obtained utilizing the nucleic acid molecules of the present invention could also be modified to affect their control characteristics. Examples of such modifications would include but are not limited to enhanced sequences as reported in Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants. Such genetic elements could be used to enhance gene expression of new and existing traits for crop improvements.

In one sub-aspect, such an analysis is conducted by determining the presence and/or identity of polymorphism(s) by one or more of the nucleic acid molecules of the present invention and more preferably one or more of the EST nucleic acid molecule or fragment thereof which are associated with a phenotype, or a predisposition to that phenotype.

Any of a variety of molecules can be used to identify such polymorphism(s). In one embodiment, one or more of the EST nucleic acid molecules (or a sub-fragment thereof) may be employed as a marker nucleic acid molecule to identify such polymorphism(s). Alternatively, such polymorphisms can be detected through the use of a marker nucleic acid molecule or a marker protein that is genetically linked to (i.e., a polynucleotide that co-segregates with) such polymorphism(s).

In an alternative embodiment, such polymorphisms can be detected through the use of a marker nucleic acid molecule that is physically linked to such polymorphism(s). For this purpose, marker nucleic acid molecules comprising a nucleotide sequence of a polynucleotide located within 1 mb of the polymorphism(s) and more preferably within 100 kb of the polymorphism(s) and most preferably within 10 kb of the polymorphism(s) can be employed.

The genomes of animals and plants naturally undergo spontaneous mutation in the course of their continuing evolution (Gusella, Ann. Rev. Biochem. 55:831-854 (1986)). A “polymorphism” is a variation or difference in the sequence of the gene or its flanking regions that arises in some of the members of a species. The variant sequence and the “original” sequence co-exist in the species' population. In some instances, such co-existence is in stable or quasi-stable equilibrium.

A polymorphism is thus said to be “allelic,” in that, due to the existence of the polymorphism, some members of a species may have the original sequence (i.e., the original “allele”) whereas other members may have the variant sequence (i.e., the variant “allele”). In the simplest case, only one variant sequence may exist and the polymorphism is thus said to be di-allelic. In other cases, the species' population may contain multiple alleles and the polymorphism is termed tri-allelic, etc. A single gene may have multiple different unrelated polymorphisms. For example, it may have a di-allelic polymorphism at one site and a multi-allelic polymorphism at another site.

The variation that defines the polymorphism may range from a single nucleotide variation to the insertion or deletion of extended regions within a gene. In some cases, the DNA sequence variations are in regions of the genome that are characterized by short tandem repeats (STRs) that include tandem di- or tri-nucleotide repeated motifs of nucleotides. Polymorphisms characterized by such tandem repeats are referred to as “variable number tandem repeat” (“VNTR”) polymorphisms. VNTRs have been used in identity analysis (Weber, U.S. Pat. No. 5,075,217; Armour et al., FEBS Lett. 307:113-115 (1992); Jones et al., Eur. J. Haematol. 39:144-147 (1987); Horn et al., PCT Patent Application WO91/14003; Jeffreys, European Patent Application 370,719; Jeffreys, U.S. Pat. No. 5,175,082; Jeffreys et al., Amer. J. Hum. Genet. 39:11-24 (1986); Jeffreys et al., Nature 316:76-79 (1985); Gray et al., Proc. R. Acad. Soc. Lond. 243:241-253 (1991); Moore et al., Genomics 10:654-660 (1991); Jeffreys et al., Anim. Genet. 18:1-15 (1987); Hillel et al., Anim. Genet. 20:145-155 (1989); Hillel et al., Genet. 124:783-789 (1990), all of which are herein incorporated by reference in their entirety).

The detection of polymorphic sites in a sample of DNA may be facilitated through the use of nucleic acid amplification methods. Such methods specifically increase the concentration of polynucleotides that span the polymorphic site, or include that site and sequences located either distal or proximal to it. Such amplified molecules can be readily detected by gel electrophoresis or other means.

The most preferred method of achieving such amplification employs the polymerase chain reaction (“PCR”) (Mullis et al., Cold Spring Harbor Symp. Quant. Biol. 51:263-273 (1986); Erlich et al., European Patent Appln. 50,424; European Patent Appln. 84,796; European Patent Application 258,017; European Patent Appln. 237,362; Mullis, European Patent Appln. 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194), using primer pairs that are capable of hybridizing to the proximal sequences that define a polymorphism in its double-stranded form.

In lieu of PCR, alternative methods, such as the “Ligase Chain Reaction” (“LCR”) may be used (Barany, Proc. Natl. Acad. Sci. (U.S.A.) 88:189-193 (1991), the entirety of which is herein incorporated by reference). LCR uses two pairs of oligonucleotide probes to exponentially amplify a specific target. The sequences of each pair of oligonucleotides is selected to permit the pair to hybridize to abutting sequences of the same strand of the target. Such hybridization forms a substrate for a template-dependent ligase. As with PCR, the resulting products thus serve as a template in subsequent cycles and an exponential amplification of the desired sequence is obtained.

LCR can be performed with oligonucleotides having the proximal and distal sequences of the same strand of a polymorphic site. In one embodiment, either oligonucleotide will be designed to include the actual polymorphic site of the polymorphism. In such an embodiment, the reaction conditions are selected such that the oligonucleotides can be ligated together only if the target molecule either contains or lacks the specific nucleotide that is complementary to the polymorphic site present on the oligonucleotide. Alternatively, the oligonucleotides may be selected such that they do not include the polymorphic site (see, Segev, PCT Application WO 90/01069, the entirety of which is herein incorporated by reference).

The “Oligonucleotide Ligation Assay” (“OLA”) may alternatively be employed (Landegren et al., Science 241:1077-1080 (1988), the entirety of which is herein incorporated by reference). The OLA protocol uses two oligonucleotides which are designed to be capable of hybridizing to abutting sequences of a single strand of a target. OLA, like LCR, is particularly suited for the detection of point mutations. Unlike LCR, however, OLA results in “linear” rather than exponential amplification of the target sequence.

Nickerson et al., have described a nucleic acid detection assay that combines attributes of PCR and OLA (Nickerson et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:8923-8927 (1990), the entirety of which is herein incorporated by reference). In this method, PCR is used to achieve the exponential amplification of target DNA, which is then detected using OLA. In addition to requiring multiple and separate, processing steps, one problem associated with such combinations is that they inherit all of the problems associated with PCR and OLA.

Schemes based on ligation of two (or more) oligonucleotides in the presence of nucleic acid having the sequence of the resulting “di-oligonucleotide”, thereby amplifying the di-oligonucleotide, are also known (Wu et al., Genomics 4:560-569 (1989), the entirety of which is herein incorporated by reference) and may be readily adapted to the purposes of the present invention.

Other known nucleic acid amplification procedures, such as allele-specific oligomers, branched DNA technology, transcription-based amplification systems, or isothermal amplification methods may also be used to amplify and analyze such polymorphisms (Malek et al., U.S. Pat. No. 5,130,238; Davey et al., European Patent Application 329,822; Schuster et al., U.S. Pat. No. 5,169,766; Miller et al., PCT Patent Application WO 89/06700; Kwoh et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:1173-1177 (1989); Gingeras et al., PCT Patent Application WO 88/10315; Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:392-396 (1992), all of which are herein incorporated by reference in their entirety).

The identification of a polymorphism can be determined in a variety of ways. By correlating the presence or absence of it in a plant with the presence or absence of a phenotype, it is possible to predict the phenotype of that plant. If a polymorphism creates or destroys a restriction endonuclease cleavage site, or if it results in the loss or insertion of DNA (e.g., a VNTR polymorphism), it will alter the size or profile of the DNA fragments that are generated by digestion with that restriction endonuclease. As such, individuals that possess a variant sequence can be distinguished from those having the original sequence by restriction fragment analysis. Polymorphisms that can be identified in this manner are termed “restriction fragment length polymorphisms” (“RFLPs”). RFLPs have been widely used in human and plant genetic analyses (Glassberg, UK Patent Application 2135774; Skolnick et al., Cytogen. Cell Genet. 32:58-67 (1982); Botstein et al., Ann. J. Hum. Genet. 32:314-331 (1980); Fischer et al., (PCT Application WO90/13668); Uhlen, PCT Application WO90/11369).

Polymorphisms can also be identified by Single Strand Conformation Polymorphism (SSCP) analysis. SSCP is a method capable of identifying most sequence variations in a single strand of DNA, typically between 150 and 250 nucleotides in length (Elles, Methods in Molecular Medicine Molecular Diagnosis of Genetic Diseases, Humana Press (1996), the entirety of which is herein incorporated by reference); Orita et al., Genomics 5:874-879 (1989), the entirety of which is herein incorporated by reference). Under denaturing conditions a single strand of DNA will adopt a conformation that is uniquely dependent on its sequence conformation. This conformation usually will be different, even if only a single base is changed. Most conformations have been reported to alter the physical configuration or size sufficiently to be detectable by electrophoresis. A number of protocols have been described for SSCP including, but not limited to, Lee et al., Anal. Biochem. 205:289-293 (1992), the entirety of which is herein incorporated by reference; Suzuki et al., Anal. Biochem. 192:82-84 (1991), the entirety of which is herein incorporated by reference; Lo et al., Nucleic Acids Research 20:1005-1009 (1992), the entirety of which is herein incorporated by reference; Sarkar et al., Genomics 13:441-443 (1992), the entirety of which is herein incorporated by reference. It is understood that one or more of the nucleic acids of the present invention, may be utilized as markers or probes to detect polymorphisms by SSCP analysis.

Polymorphisms may also be found using a DNA fingerprinting technique called amplified fragment length polymorphism (AFLP), which is based on the selective PCR amplification of restriction fragments from a total digest of genomic DNA to profile that DNA (Vos et al., Nucleic Acids Res. 23:4407-4414 (1995), the entirety of which is herein incorporated by reference). This method allows for the specific co-amplification of high numbers of restriction fragments, which can be visualized by PCR without knowledge of the nucleic acid sequence.

AFLP employs basically three steps. Initially, a sample of genomic DNA is cut with restriction enzymes and oligonucleotide adapters are ligated to the restriction fragments of the DNA. The restriction fragments are then amplified using PCR by using the adapter and restriction sequence as target sites for primer annealing. The selective amplification is achieved by the use of primers that extend into the restriction fragments, amplifying only those fragments in which the primer extensions match the nucleotide flanking the restriction sites. These amplified fragments are then visualized on a denaturing polyacrylamide gel.

AFLP analysis has been performed on Salix (Beismann et al., Mol. Ecol. 6:989-993 (1997), the entirety of which is herein incorporated by reference), Acinetobacter (Janssen et al., Int. J. Syst. Bacteriol. 47:1179-1187 (1997), the entirety of which is herein incorporated by reference), Aeromonas popoffi (Huys et al., Int. J. Syst. Bacteriol. 47:1165-1171 (1997), the entirety of which is herein incorporated by reference), rice (McCouch et al., Plant Mol. Biol. 35:89-99 (1997), the entirety of which is herein incorporated by reference; Nandi et al., Mol. Gen. Genet. 255:1-8 (1997), the entirety of which is herein incorporated by reference; Cho et al., Genome 39:373-378 (1996), the entirety of which is herein incorporated by reference), barley (Hordeum vulgare) (Simons et al., Genomics 44:61-70 (1997), the entirety of which is herein incorporated by reference; Waugh et al., Mol. Gen. Genet. 255:311-321 (1997), the entirety of which is herein incorporated by reference; Qi et al., Mol. Gen. Genet. 254:330-336 (1997), the entirety of which is herein incorporated by reference; Becker et al., Mol. Gen. Genet. 249:65-73 (1995), the entirety of which is herein incorporated by reference), potato (Van der Voort et al., Mol. Gen. Genet. 255:438-447 (1997), the entirety of which is herein incorporated by reference; Meksem et al., Mol. Gen. Genet. 249:74-81 (1995), the entirety of which is herein incorporated by reference), Phytophthora infestans (Van der Lee et al., Fungal Genet. Biol. 21:278-291 (1997), the entirety of which is herein incorporated by reference), Bacillus anthracis (Keim et al., J. Bacteriol. 179:818-824 (1997), the entirety of which is herein incorporated by reference), Astragalus cremnophylax (Travis et al., Mol. Ecol. 5:735-745 (1996), the entirety of which is herein incorporated by reference), Arabidopsis (Cnops et al., Mol. Gen. Genet. 253:32-41 (1996), the entirety of which is herein incorporated by reference), Escherichia coli (Lin et al., Nucleic Acids Res. 24:3649-3650 (1996), the entirety of which is herein incorporated by reference), Aeromonas (Huys et al., Int. J. Syst. Bacteriol. 46:572-580 (1996), the entirety of which is herein incorporated by reference), nematode (Folkertsma et al., Mol. Plant Microbe Interact. 9:47-54 (1996), the entirety of which is herein incorporated by reference), tomato (Thomas et al., Plant J. 8:785-794 (1995), the entirety of which is herein incorporated by reference) and human (Latorra et al., PCR Methods Appl. 3:351-358 (1994), the entirety of which is herein incorporated by reference). AFLP analysis has also been used for fingerprinting mRNA (Money et al., Nucleic Acids Res. 24:2616-2617 (1996), the entirety of which is herein incorporated by reference; Bachem et al., Plant J. 9:745-753 (1996), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acids of the present invention, may be utilized as markers or probes to detect polymorphisms by AFLP analysis or for fingerprinting RNA.

Polymorphisms may also be found using random amplified polymorphic DNA (RAPD) (Williams et al., Nucl. Acids Res. 18:6531-6535 (1990), the entirety of which is herein incorporated by reference) and cleaveable amplified polymorphic sequences (CAPS) (Lyamichev et al., Science 260:778-783 (1993), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules of the present invention, may be utilized as markers or probes to detect polymorphisms by RAPD or CAPS analysis.

Through genetic mapping, a fine scale linkage map can be developed using DNA markers and, then, a genomic DNA library of large-sized fragments can be screened with molecular markers linked to the desired trait. Molecular markers are advantageous for agronomic traits that are otherwise difficult to tag, such as resistance to pathogens, insects and nematodes, tolerance to abiotic stress, quality parameters and quantitative traits such as high yield potential.

The essential requirements for marker-assisted selection in a plant breeding program are: (1) the marker(s) should co-segregate or be closely linked with the desired trait; (2) an efficient means of screening large populations for the molecular marker(s) should be available; and (3) the screening technique should have high reproducibility across laboratories and preferably be economical to use and be user-friendly.

The genetic linkage of marker molecules can be established by a gene mapping model such as, without limitation, the flanking marker model reported by Lander and Botstein, Genetics 121:185-199 (1989) and the interval mapping, based on maximum likelihood methods described by Lander and Botstein, Genetics 121:185-199 (1989) and implemented in the software package MAPMAKER/QTL (Lincoln and Lander, Mapping Genes Controlling Quantitative Traits Using MAPMAKER/QTL, Whitehead Institute for Biomedical Research, Massachusetts, (1990). Additional software includes Qgene, Version 2.23 (1996), Department of Plant Breeding and Biometry, 266 Emerson Hall, Cornell University, Ithaca, N.Y., the manual of which is herein incorporated by reference in its entirety). Use of Qgene software is a particularly preferred approach.

A maximum likelihood estimate (MLE) for the presence of a marker is calculated, together with an MLE assuming no QTL effect, to avoid false positives. A log₁₀ of an odds ratio (LOD) is then calculated as: LOD=log₁₀ (MLE for the presence of a QTL/MLE given no linked QTL).

The LOD score essentially indicates how much more likely the data are to have arisen assuming the presence of a QTL than in its absence. The LOD threshold value for avoiding a false positive with a given confidence, say 95%, depends on the number of markers and the length of the genome. Graphs indicating LOD thresholds are set forth in Lander and Botstein, Genetics 121:185-199 (1989) the entirety of which is herein incorporated by reference and further described by Arús and Moreno-González, Plant Breeding, Hayward et al., (eds.) Chapman & Hall, London, pp. 314-331 (1993), the entirety of which is herein incorporated by reference.

Additional models can be used. Many modifications and alternative approaches to interval mapping have been reported, including the use non-parametric methods (Kruglyak and Lander, Genetics 139:1421-1428 (1995), the entirety of which is herein incorporated by reference). Multiple regression methods or models can be also be used, in which the trait is regressed on a large number of markers (Jansen, Biometrics in Plant Breeding, van Oijen and Jansen (eds.), Proceedings of the Ninth Meeting of the Eucarpia Section Biometrics in Plant Breeding, The Netherlands, pp. 116-124 (1994); Weber and Wricke, Advances in Plant Breeding, Blackwell, Berlin, 16 (1994), both of which is herein incorporated by reference in their entirety). Procedures combining interval mapping with regression analysis, whereby the phenotype is regressed onto a single putative QTL at a given marker interval and at the same time onto a number of markers that serve as ‘cofactors,’ have been reported by Jansen and Stam, Genetics 136:1447-1455 (1994), the entirety of which is herein incorporated by reference and Zeng, Genetics 136:1457-1468 (1994) the entirety of which is herein incorporated by reference. Generally, the use of cofactors reduces the bias and sampling error of the estimated QTL positions (Utz and Melchinger, Biometrics in Plant Breeding, van Oijen and Jansen (eds.) Proceedings of the Ninth Meeting of the Eucarpia Section Biometrics in Plant Breeding, The Netherlands, pp. 195-204 (1994), the entirety of which is herein incorporated by reference, thereby improving the precision and efficiency of QTL mapping (Zeng, Genetics 136:1457-1468 (1994)). These models can be extended to multi-environment experiments to analyze genotype-environment interactions (Jansen et al., Theo. Appl. Genet. 91:33-37 (1995), the entirety of which is herein incorporated by reference).

Selection of an appropriate mapping populations is important to map construction. The choice of appropriate mapping population depends on the type of marker systems employed (Tanksley et al., Molecular mapping plant chromosomes. Chromosome structure and function: Impact of new concepts, Gustafson and Appels (eds.), Plenum Press, New York, pp 157-173 (1988), the entirety of which is herein incorporated by reference). Consideration must be given to the source of parents (adapted vs. exotic) used in the mapping population. Chromosome pairing and recombination rates can be severely disturbed (suppressed) in wide crosses (adapted×exotic) and generally yield greatly reduced linkage distances. Wide crosses will usually provide segregating populations with a relatively large array of polymorphisms when compared to progeny in a narrow cross (adapted×adapted).

An F₂ population is the first generation of selfing after the hybrid seed is produced. Usually a single F₁ plant is selfed to generate a population segregating for all the genes in Mendelian (1:2:1) fashion. Maximum genetic information is obtained from a completely classified F₂ population using a codominant marker system (Mather, Measurement of Linkage in Heredity, Methuen and Co., (1938), the entirety of which is herein incorporated by reference). In the case of dominant markers, progeny tests (e.g. F₃, BCF₂) are required to identify the heterozygotes, thus making it equivalent to a completely classified F₂ population. However, this procedure is often prohibitive because of the cost and time involved in progeny testing. Progeny testing of F₂ individuals is often used in map construction where phenotypes do not consistently reflect genotype (e.g. disease resistance) or where trait expression is controlled by a QTL. Segregation data from progeny test populations (e.g. F₃ or BCF₂) can be used in map construction. Marker-assisted selection can then be applied to cross progeny based on marker-trait map associations (F₂, F₃), where linkage groups have not been completely disassociated by recombination events (i.e., maximum disequillibrium).

Recombinant inbred lines (RIL) (genetically related lines; usually >F₅, developed from continuously selfing F₂ lines towards homozygosity) can be used as a mapping population. Information obtained from dominant markers can be maximized by using RIL because all loci are homozygous or nearly so. Under conditions of tight linkage (i.e., about <10% recombination), dominant and co-dominant markers evaluated in RIL populations provide more information per individual than either marker type in backcross populations (Reiter et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992), the entirety of which is herein incorporated by reference). However, as the distance between markers becomes larger (i.e., loci become more independent), the information in RIL populations decreases dramatically when compared to codominant markers.

Backcross populations (e.g., generated from a cross between a successful variety (recurrent parent) and another variety (donor parent) carrying a trait not present in the former) can be utilized as a mapping population. A series of backcrosses to the recurrent parent can be made to recover most of its desirable traits. Thus a population is created consisting of individuals nearly like the recurrent parent but each individual carries varying amounts or mosaic of genomic regions from the donor parent. Backcross populations can be useful for mapping dominant markers if all loci in the recurrent parent are homozygous and the donor and recurrent parent have contrasting polymorphic marker alleles (Reiter et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992)). Information obtained from backcross populations using either codominant or dominant markers is less than that obtained from F₂ populations because one, rather than two, recombinant gametes are sampled per plant. Backcross populations, however, are more informative (at low marker saturation) when compared to RILs as the distance between linked loci increases in RIL populations (i.e. about 15% recombination). Increased recombination can be beneficial for resolution of tight linkages, but may be undesirable in the construction of maps with low marker saturation.

Near-isogenic lines (NIL) created by many backcrosses to produce an array of individuals that are nearly identical in genetic composition except for the trait or genomic region under interrogation can be used as a mapping population. In mapping with NILs, only a portion of the polymorphic loci are expected to map to a selected region.

Bulk segregant analysis (BSA) is a method developed for the rapid identification of linkage between markers and traits of interest (Michelmore et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:9828-9832 (1991), the entirety of which is herein incorporated by reference). In BSA, two bulked DNA samples are drawn from a segregating population originating from a single cross. These bulks contain individuals that are identical for a particular trait (resistant or susceptible to particular disease) or genomic region but arbitrary at unlinked regions (i.e. heterozygous). Regions unlinked to the target region will not differ between the bulked samples of many individuals in BSA.

It is understood that one or more of the nucleic acid molecules of the present invention may be used as molecular markers. It is also understood that one or more of the protein molecules of the present invention may be used as molecular markers.

In accordance with this aspect of the present invention, a sample nucleic acid is obtained from plants cells or tissues. Any source of nucleic acid may be used. Preferably, the nucleic acid is genomic DNA. The nucleic acid is subjected to restriction endonuclease digestion. For example, one or more nucleic acid molecule or fragment thereof of the present invention can be used as a probe in accordance with the above-described polymorphic methods. The polymorphism obtained in this approach can then be cloned to identify the mutation at the coding region which alters the protein's structure or regulatory region of the gene which affects its expression level.

In an aspect of the present invention, one or more of the nucleic molecules of the present invention are used to determine the level (i.e., the concentration of mRNA in a sample, etc.) in a plant (preferably maize or soybean) or pattern (i.e., the kinetics of expression, rate of decomposition, stability profile, etc.) of the expression of a protein encoded in part or whole by one or more of the nucleic acid molecule of the present invention (collectively, the “Expression Response” of a cell or tissue). As used herein, the Expression Response manifested by a cell or tissue is said to be “altered” if it differs from the Expression Response of cells or tissues of plants not exhibiting the phenotype. To determine whether a Expression Response is altered, the Expression Response manifested by the cell or tissue of the plant exhibiting the phenotype is compared with that of a similar cell or tissue sample of a plant not exhibiting the phenotype. As will be appreciated, it is not necessary to re-determine the Expression Response of the cell or tissue sample of plants not exhibiting the phenotype each time such a comparison is made; rather, the Expression Response of a particular plant may be compared with previously obtained values of normal plants. As used herein, the phenotype of the organism is any of one or more characteristics of an organism (e.g. disease resistance, pest tolerance, environmental tolerance such as tolerance to abiotic stress, male sterility, quality improvement or yield etc.). A change in genotype or phenotype may be transient or permanent. Also as used herein, a tissue sample is any sample that comprises more than one cell. In a preferred aspect, a tissue sample comprises cells that share a common characteristic (e.g. derived from root, seed, flower, leaf, stem or pollen etc.).

In one aspect of the present invention, an evaluation can be conducted to determine whether a particular mRNA molecule is present. One or more of the nucleic acid molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention are utilized to detect the presence or quantity of the mRNA species. Such molecules are then incubated with cell or tissue extracts of a plant under conditions sufficient to permit nucleic acid hybridization. The detection of double-stranded probe-mRNA hybrid molecules is indicative of the presence of the mRNA; the amount of such hybrid formed is proportional to the amount of mRNA. Thus, such probes may be used to ascertain the level and extent of the mRNA production in a plant's cells or tissues. Such nucleic acid hybridization may be conducted under quantitative conditions (thereby providing a numerical value of the amount of the mRNA present). Alternatively, the assay may be conducted as a qualitative assay that indicates either that the mRNA is present, or that its level exceeds a user set, predefined value.

A principle of in situ hybridization is that a labeled, single-stranded nucleic acid probe will hybridize to a complementary strand of cellular DNA or RNA and, under the appropriate conditions, these molecules will form a stable hybrid. When nucleic acid hybridization is combined with histological techniques, specific DNA or RNA sequences can be identified within a single cell. An advantage of in situ hybridization over more conventional techniques for the detection of nucleic acids is that it allows an investigator to determine the precise spatial population (Angerer et al., Dev. Biol. 101:477-484 (1984), the entirety of which is herein incorporated by reference; Angerer et al., Dev. Biol. 112:157-166 (1985), the entirety of which is herein incorporated by reference; Dixon et al., EMBO J. 10:1317-1324 (1991), the entirety of which is herein incorporated by reference). In situ hybridization may be used to measure the steady-state level of RNA accumulation. It is a sensitive technique and RNA sequences present in as few as 5-10 copies per cell can be detected (Hardin et al., J. Mol. Biol. 202:417-431 (1989), the entirety of which is herein incorporated by reference). A number of protocols have been devised for in situ hybridization, each with tissue preparation, hybridization and washing conditions (Meyerowitz, Plant Mol. Biol. Rep. 5:242-250 (1987), the entirety of which is herein incorporated by reference; Cox and Goldberg, In: Plant Molecular Biology: A Practical Approach, Shaw (ed.), pp 1-35, IRL Press, Oxford (1988), the entirety of which is herein incorporated by reference; Raikhel et al., In situ RNA hybridization in plant tissues, In: Plant Molecular Biology Manual, vol. B9: 1-32, Kluwer Academic Publisher, Dordrecht, Belgium (1989), the entirety of which is herein incorporated by reference).

In situ hybridization also allows for the localization of proteins within a tissue or cell (Wilkinson, In Situ Hybridization, Oxford University Press, Oxford (1992), the entirety of which is herein incorporated by reference; Langdale, In Situ Hybridization In: The Maize Handbook Freeling and Walbot (eds.), pp 165-179, Springer-Verlag, New York (1994), the entirety of which is herein incorporated by reference). It is understood that one or more of the molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention or one or more of the antibodies of the present invention may be utilized to detect the level or pattern of a methionine pathway protein or mRNA thereof by in situ hybridization.

Fluorescent in situ hybridization allows the localization of a particular DNA sequence along a chromosome which is useful, among other uses, for gene mapping, following chromosomes in hybrid lines or detecting chromosomes with translocations, transversions or deletions. In situ hybridization has been used to identify chromosomes in several plant species (Griffor et al., Plant Mol. Biol. 17:101-109 (1991), the entirety of which is herein incorporated by reference; Gustafson et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:1899-1902 (1990), herein incorporated by reference; Mukai and Gill, Genome 34:448-452 (1991), the entirety of which is herein incorporated by reference; Schwarzacher and Heslop-Harrison, Genome 34:317-323 (1991); Wang et al., Jpn. J. Genet. 66:313-316 (1991), the entirety of which is herein incorporated by reference; Parra and Windle, Nature Genetics 5:17-21 (1993), the entirety of which is herein incorporated by reference). It is understood that the nucleic acid molecules of the present invention may be used as probes or markers to localize sequences along a chromosome.

Another method to localize the expression of a molecule is tissue printing. Tissue printing provides a way to screen, at the same time on the same membrane many tissue sections from different plants or different developmental stages. Tissue-printing procedures utilize films designed to immobilize proteins and nucleic acids. In essence, a freshly cut section of a tissue is pressed gently onto nitrocellulose paper, nylon membrane or polyvinylidene difluoride membrane. Such membranes are commercially available (e.g. Millipore, Bedford, Mass. U.S.A.). The contents of the cut cell transfer onto the membrane and the contents and are immobilized to the membrane. The immobilized contents form a latent print that can be visualized with appropriate probes. When a plant tissue print is made on nitrocellulose paper, the cell walls leave a physical print that makes the anatomy visible without further treatment (Varner and Taylor, Plant Physiol. 91:31-33 (1989), the entirety of which is herein incorporated by reference).

Tissue printing on substrate films is described by Daoust, Exp. Cell Res. 12:203-211 (1957), the entirety of which is herein incorporated by reference, who detected amylase, protease, ribonuclease and deoxyribonuclease in animal tissues using starch, gelatin and agar films. These techniques can be applied to plant tissues (Yomo and Taylor, Planta 112:35-43 (1973); the entirety of which is herein incorporated by reference; Harris and Chrispeels, Plant Physiol. 56:292-299 (1975), the entirety of which is herein incorporated by reference). Advances in membrane technology have increased the range of applications of Daoust's tissue-printing techniques allowing (Cassab and Varner, J. Cell. Biol. 105:2581-2588 (1987), the entirety of which is herein incorporated by reference) the histochemical localization of various plant enzymes and deoxyribonuclease on nitrocellulose paper and nylon (Spruce et al., Phytochemistry 26:2901-2903 (1987), the entirety of which is herein incorporated by reference; Barres et al., Neuron 5:527-544 (1990), the entirety of which is herein incorporated by reference; Reid and Pont-Lezica, Tissue Printing: Tools for the Study of Anatomy, Histochemistry and Gene Expression, Academic Press, New York, N.Y. (1992), the entirety of which is herein incorporated by reference; Reid et al., Plant Physiol. 93:160-165 (1990), the entirety of which is herein incorporated by reference; Ye et al., Plant J. 1:175-183 (1991), the entirety of which is herein incorporated by reference).

It is understood that one or more of the molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention or one or more of the antibodies of the present invention may be utilized to detect the presence or quantity of a methionine pathway protein by tissue printing.

Further it is also understood that any of the nucleic acid molecules of the present invention may be used as marker nucleic acids and or probes in connection with methods that require probes or marker nucleic acids. As used herein, a probe is an agent that is utilized to determine an attribute or feature (e.g. presence or absence, location, correlation, etc.) of a molecule, cell, tissue or plant. As used herein, a marker nucleic acid is a nucleic acid molecule that is utilized to determine an attribute or feature (e.g., presence or absence, location, correlation, etc.) or a molecule, cell, tissue or plant.

A microarray-based method for high-throughput monitoring of plant gene expression may be utilized to measure gene-specific hybridization targets. This ‘chip’-based approach involves using microarrays of nucleic acid molecules as gene-specific hybridization targets to quantitatively measure expression of the corresponding plant genes (Schena et al., Science 270:467-470 (1995), the entirety of which is herein incorporated by reference; Shalon, Ph.D. Thesis, Stanford University (1996), the entirety of which is herein incorporated by reference). Every nucleotide in a large sequence can be queried at the same time. Hybridization can be used to efficiently analyze nucleotide sequences.

Several microarray methods have been described. One method compares the sequences to be analyzed by hybridization to a set of oligonucleotides representing all possible subsequences (Bains and Smith, J. Theor. Biol. 135:303-307 (1989), the entirety of which is herein incorporated by reference). A second method hybridizes the sample to an array of oligonucleotide or cDNA molecules. An array consisting of oligonucleotides complementary to subsequences of a target sequence can be used to determine the identity of a target sequence, measure its amount and detect differences between the target and a reference sequence. Nucleic acid molecules microarrays may also be screened with protein molecules or fragments thereof to determine nucleic acid molecules that specifically bind protein molecules or fragments thereof.

The microarray approach may be used with polypeptide targets (U.S. Pat. No. 5,445,934; U.S. Pat. No. 5,143,854; U.S. Pat. No. 5,079,600; U.S. Pat. No. 4,923,901, all of which are herein incorporated by reference in their entirety). Essentially, polypeptides are synthesized on a substrate (microarray) and these polypeptides can be screened with either protein molecules or fragments thereof or nucleic acid molecules in order to screen for either protein molecules or fragments thereof or nucleic acid molecules that specifically bind the target polypeptides. (Fodor et al., Science 251:767-773 (1991), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules or protein or fragments thereof of the present invention may be utilized in a microarray based method.

In a preferred embodiment of the present invention microarrays may be prepared that comprise nucleic acid molecules where such nucleic acid molecules encode at least one, preferably at least two, more preferably at least three, even more preferably at least four, five six or seven methionine pathway enzymes. In a preferred embodiment the nucleic acid molecules are selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean methionine adenosyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean S-adenosylmethionine decarboxylase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-lyase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean cystathionine γ-lyase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof.

Site directed mutagenesis may be utilized to modify nucleic acid sequences, particularly as it is a technique that allows one or more of the amino acids encoded by a nucleic acid molecule to be altered (e.g. a threonine to be replaced by a methionine). Three basic methods for site directed mutagenesis are often employed. These are cassette mutagenesis (Wells et al., Gene 34:315-323 (1985), the entirety of which is herein incorporated by reference), primer extension (Gilliam et al., Gene 12:129-137 (1980), the entirety of which is herein incorporated by reference; Zoller and Smith, Methods Enzymol. 100:468-500 (1983), the entirety of which is herein incorporated by reference; Dalbadie-McFarland et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:6409-6413 (1982), the entirety of which is herein incorporated by reference) and methods based upon PCR (Scharf et al., Science 233:1076-1078 (1986), the entirety of which is herein incorporated by reference; Higuchi et al., Nucleic Acids Res. 16:7351-7367 (1988), the entirety of which is herein incorporated by reference). Site directed mutagenesis approaches are also described in European Patent 0 385 962, the entirety of which is herein incorporated by reference; European Patent 0 359 472, the entirety of which is herein incorporated by reference; and PCT Patent Application WO 93/07278, the entirety of which is herein incorporated by reference.

Site directed mutagenesis strategies have been applied to plants for both in vitro as well as in vivo site directed mutagenesis (Lanz et al., J. Biol. Chem. 266:9971-9976 (1991), the entirety of which is herein incorporated by reference; Kovgan and Zhdanov, Biotekhnologiya 5:148-154, No. 207160n, Chemical Abstracts 110:225 (1989), the entirety of which is herein incorporated by reference; Ge et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:4037-4041 (1989), the entirety of which is herein incorporated by reference; Zhu et al., J. Biol. Chem. 271:18494-18498 (1996), the entirety of which is herein incorporated by reference; Chu et al., Biochemistry 33:6150-6157 (1994), the entirety of which is herein incorporated by reference; Small et al., EMBO J. 11:1291-1296 (1992), the entirety of which is herein incorporated by reference; Cho et al., Mol. Biotechnol. 8:13-16 (1997), the entirety of which is herein incorporated by reference; Kita et al., J. Biol. Chem. 271:26529-26535 (1996), the entirety of which is herein incorporated by reference, Jin et al., Mol. Microbiol. 7:555-562 (1993), the entirety of which is herein incorporated by reference; Hatfield and Vierstra, J. Biol. Chem. 267:14799-14803 (1992), the entirety of which is herein incorporated by reference; Zhao et al., Biochemistry 31:5093-5099 (1992), the entirety of which is herein incorporated by reference).

Any of the nucleic acid molecules of the present invention may either be modified by site directed mutagenesis or used as, for example, nucleic acid molecules that are used to target other nucleic acid molecules for modification. It is understood that mutants with more than one altered nucleotide can be constructed using techniques that practitioners are familiar with such as isolating restriction fragments and ligating such fragments into an expression vector (see, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989)).

Sequence-specific DNA-binding proteins play a role in the regulation of transcription. The isolation of recombinant cDNAs encoding these proteins facilitates the biochemical analysis of their structural and functional properties. Genes encoding such DNA-binding proteins have been isolated using classical genetics (Vollbrecht et al., Nature 350: 241-243 (1991), the entirety of which is herein incorporated by reference) and molecular biochemical approaches, including the screening of recombinant cDNA libraries with antibodies (Landschulz et al., Genes Dev. 2:786-800 (1988), the entirety of which is herein incorporated by reference) or DNA probes (Bodner et al., Cell 55:505-518 (1988), the entirety of which is herein incorporated by reference). In addition, an in situ screening procedure has been used and has facilitated the isolation of sequence-specific DNA-binding proteins from various plant species (Gilmartin et al., Plant Cell 4:839-849 (1992), the entirety of which is herein incorporated by reference; Schindler et al., EMBO J. 11:1261-1273 (1992), the entirety of which is herein incorporated by reference). An in situ screening protocol does not require the purification of the protein of interest (Vinson et al., Genes Dev. 2:801-806 (1988), the entirety of which is herein incorporated by reference; Singh et al., Cell 52:415-423 (1988), the entirety of which is herein incorporated by reference).

Two steps may be employed to characterize DNA-protein interactions. The first is to identify promoter fragments that interact with DNA-binding proteins, to titrate binding activity, to determine the specificity of binding and to determine whether a given DNA-binding activity can interact with related DNA sequences (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2^(nd) edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)). Electrophoretic mobility-shift assay is a widely used assay. The assay provides a rapid and sensitive method for detecting DNA-binding proteins based on the observation that the mobility of a DNA fragment through a nondenaturing, low-ionic strength polyacrylamide gel is retarded upon association with a DNA-binding protein (Fried and Crother, Nucleic Acids Res. 9:6505-6525 (1981), the entirety of which is herein incorporated by reference). When one or more specific binding activities have been identified, the exact sequence of the DNA bound by the protein may be determined. Several procedures for characterizing protein/DNA-binding sites are used, including methylation and ethylation interference assays (Maxam and Gilbert, Methods Enzymol. 65:499-560 (1980), the entirety of which is herein incorporated by reference; Wissman and Hillen, Methods Enzymol. 208:365-379 (1991), the entirety of which is herein incorporated by reference), footprinting techniques employing DNase I (Galas and Schmitz, Nucleic Acids Res. 5:3157-3170 (1978), the entirety of which is herein incorporated by reference), 1,10-phenanthroline-copper ion methods (Sigman et al., Methods Enzymol. 208:414-433 (1991), the entirety of which is herein incorporated by reference) and hydroxyl radicals methods (Dixon et al., Methods Enzymol. 208:414-433 (1991), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules of the present invention may be utilized to identify a protein or fragment thereof that specifically binds to a nucleic acid molecule of the present invention. It is also understood that one or more of the protein molecules or fragments thereof of the present invention may be utilized to identify a nucleic acid molecule that specifically binds to it.

A two-hybrid system is based on the fact that many cellular functions are carried out by proteins, such as transcription factors, that interact (physically) with one another. Two-hybrid systems have been used to probe the function of new proteins (Chien et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:9578-9582 (1991) the entirety of which is herein incorporated by reference; Durfee et al., Genes Dev. 7:555-569 (1993) the entirety of which is herein incorporated by reference; Choi et al., Cell 78:499-512 (1994), the entirety of which is herein incorporated by reference; Kranz et al., Genes Dev. 8:313-327 (1994), the entirety of which is herein incorporated by reference).

Interaction mating techniques have facilitated a number of two-hybrid studies of protein-protein interaction. Interaction mating has been used to examine interactions between small sets of tens of proteins (Finley and Brent, Proc. Natl. Acad. Sci. (U.S.A.) 91:12098-12984 (1994), the entirety of which is herein incorporated by reference), larger sets of hundreds of proteins (Bendixen et al., Nucl. Acids Res. 22:1778-1779 (1994), the entirety of which is herein incorporated by reference) and to comprehensively map proteins encoded by a small genome (Bartel et al., Nature Genetics 12:72-77 (1996), the entirety of which is herein incorporated by reference). This technique utilizes proteins fused to the DNA-binding domain and proteins fused to the activation domain. They are expressed in two different haploid yeast strains of opposite mating type and the strains are mated to determine if the two proteins interact. Mating occurs when haploid yeast strains come into contact and result in the fusion of the two haploids into a diploid yeast strain. An interaction can be determined by the activation of a two-hybrid reporter gene in the diploid strain. An advantage of this technique is that it reduces the number of yeast transformations needed to test individual interactions. It is understood that the protein-protein interactions of protein or fragments thereof of the present invention may be investigated using the two-hybrid system and that any of the nucleic acid molecules of the present invention that encode such proteins or fragments thereof may be used to transform yeast in the two-hybrid system.

(a) Plant Constructs and Plant Transformants

One or more of the nucleic acid molecules of the present invention may be used in plant transformation or transfection. Exogenous genetic material may be transferred into a plant cell and the plant cell regenerated into a whole, fertile or sterile plant. Exogenous genetic material is any genetic material, whether naturally occurring or otherwise, from any source that is capable of being inserted into any organism. Such genetic material may be transferred into either monocotyledons and dicotyledons including, but not limited to maize (pp 63-69), soybean (pp 50-60), Arabidopsis (p 45), phaseolus (pp 47-49), peanut (pp 49-50), alfalfa (p 60), wheat (pp 69-71), rice (pp 72-79), oat (pp 80-81), sorghum (p 83), rye (p 84), tritordeum (p 84), millet (p 85), fescue (p 85), perennial ryegrass (p 86), sugarcane (p 87), cranberry (p 110), papaya (pp 101-102), banana (p 103), banana (p 103), muskmelon (p 104), apple (p 104), cucumber (p 105), dendrobium (p 109), gladiolus (p 110), chrysanthemum (p 110), liliacea (p 111), cotton (pp 113-114), eucalyptus (p 115), sunflower (p 118), canola (p 118), turfgrass (p 121), sugarbeet (p 122), coffee (p 122) and dioscorea (p 122), (Christou, In: Particle Bombardment for Genetic Engineering of Plants, Biotechnology Intelligence Unit. Academic Press, San Diego, Calif. (1996), the entirety of which is herein incorporated by reference).

Transfer of a nucleic acid that encodes for a protein can result in overexpression of that protein in a transformed cell or transgenic plant. One or more of the proteins or fragments thereof encoded by nucleic acid molecules of the present invention may be overexpressed in a transformed cell or transformed plant. Particularly, any of the methionine pathway proteins or fragments thereof may be overexpressed in a transformed cell or transgenic plant. Such overexpression may be the result of transient or stable transfer of the exogenous genetic material.

Exogenous genetic material may be transferred into a plant cell and the plant cell by the use of a DNA vector or construct designed for such a purpose. Design of such a vector is generally within the skill of the art (See, Plant Molecular Biology: A Laboratory Manual, Clark (ed.), Springier, N.Y. (1997), the entirety of which is herein incorporated by reference).

A construct or vector may include a plant promoter to express the protein or protein fragment of choice. A number of promoters which are active in plant cells have been described in the literature. These include the nopaline synthase (NOS) promoter (Ebert et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:5745-5749 (1987), the entirety of which is herein incorporated by reference), the octopine synthase (OCS) promoter (which are carried on tumor-inducing plasmids of Agrobacterium tumefaciens), the caulimovirus promoters such as the cauliflower mosaic virus (CaMV) 19S promoter (Lawton et al., Plant Mol. Biol. 9:315-324 (1987), the entirety of which is herein incorporated by reference) and the CAMV 35S promoter (Odell et al., Nature 313:810-812 (1985), the entirety of which is herein incorporated by reference), the figwort mosaic virus 35S-promoter, the light-inducible promoter from the small subunit of ribulose-1,5-bis-phosphate carboxylase (ssRUBISCO), the Adh promoter (Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:6624-6628 (1987), the entirety of which is herein incorporated by reference), the sucrose synthase promoter (Yang et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:4144-4148 (1990), the entirety of which is herein incorporated by reference), the R gene complex promoter (Chandler et al., The Plant Cell 1:1175-1183 (1989), the entirety of which is herein incorporated by reference) and the chlorophyll a/b binding protein gene promoter, etc. These promoters have been used to create DNA constructs which have been expressed in plants; see, e.g., PCT publication WO 84/02913, herein incorporated by reference in its entirety.

Promoters which are known or are found to cause transcription of DNA in plant cells can be used in the present invention. Such promoters may be obtained from a variety of sources such as plants and plant viruses. It is preferred that the particular promoter selected should be capable of causing sufficient expression to result in the production of an effective amount of the methionine pathway protein to cause the desired phenotype. In addition to promoters that are known to cause transcription of DNA in plant cells, other promoters may be identified for use in the current invention by screening a plant cDNA library for genes which are selectively or preferably expressed in the target tissues or cells.

For the purpose of expression in source tissues of the plant, such as the leaf, seed, root or stem, it is preferred that the promoters utilized in the present invention have relatively high expression in these specific tissues. For this purpose, one may choose from a number of promoters for genes with tissue- or cell-specific or -enhanced expression. Examples of such promoters reported in the literature include the chloroplast glutamine synthetase GS2 promoter from pea (Edwards et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:3459-3463 (1990), herein incorporated by reference in its entirety), the chloroplast fructose-1,6-biphosphatase (FBPase) promoter from wheat (Lloyd et al., Mol. Gen. Genet. 225:209-216 (1991), herein incorporated by reference in its entirety), the nuclear photosynthetic ST-LS1 promoter from potato (Stockhaus et al., EMBO J. 8:2445-2451 (1989), herein incorporated by reference in its entirety), the serine/threonine kinase (PAL) promoter and the glucoamylase (CHS) promoter from Arabidopsis thaliana. Also reported to be active in photosynthetically active tissues are the ribulose-1,5-bisphosphate carboxylase (RbcS) promoter from eastern larch (Larix laricina), the promoter for the cab gene, cab6, from pine (Yamamoto et al., Plant Cell Physiol. 35:773-778 (1994), herein incorporated by reference in its entirety), the promoter for the Cab-1 gene from wheat (Fejes et al., Plant Mol. Biol. 15:921-932 (1990), herein incorporated by reference in its entirety), the promoter for the CAB-1 gene from spinach (Lubberstedt et al., Plant Physiol. 104:997-1006 (1994), herein incorporated by reference in its entirety), the promoter for the cab1R gene from rice (Luan et al., Plant Cell. 4:971-981 (1992), the entirety of which is herein incorporated by reference), the pyruvate, orthophosphate dikinase (PPDK) promoter from maize (Matsuoka et al., Proc. Natl. Acad. Sci. (U.S.A.) 90: 9586-9590 (1993), herein incorporated by reference in its entirety), the promoter for the tobacco Lhcb1*2 gene (Cerdan et al., Plant Mol. Biol. 33:245-255 (1997), herein incorporated by reference in its entirety), the Arabidopsis thaliana SUC2 sucrose-H+ symporter promoter (Truernit et al., Planta. 196:564-570 (1995), herein incorporated by reference in its entirety) and the promoter for the thylakoid membrane proteins from spinach (psaD, psaF, psaE, PC, FNR, atpC, atpD, cab, rbcS). Other promoters for the chlorophyll a/b-binding proteins may also be utilized in the present invention, such as the promoters for LhcB gene and PsbP gene from white mustard (Sinapis alba; Kretsch et al., Plant Mol. Biol. 28:219-229 (1995), the entirety of which is herein incorporated by reference).

For the purpose of expression in sink tissues of the plant, such as the tuber of the potato plant, the fruit of tomato, or the seed of maize, wheat, rice and barley, it is preferred that the promoters utilized in the present invention have relatively high expression in these specific tissues. A number of promoters for genes with tuber-specific or -enhanced expression are known, including the class I patatin promoter (Bevan et al., EMBO J. 8:1899-1906 (1986); Jefferson et al., Plant Mol. Biol. 14:995-1006 (1990), both of which are herein incorporated by reference in its entirety), the promoter for the potato tuber ADPGPP genes, both the large and small subunits, the sucrose synthase promoter (Salanoubat and Belliard, Gene. 60:47-56 (1987), Salanoubat and Belliard, Gene. 84:181-185 (1989), both of which are incorporated by reference in their entirety), the promoter for the major tuber proteins including the 22 kd protein complexes and proteinase inhibitors (Hannapel, Plant Physiol. 101:703-704 (1993), herein incorporated by reference in its entirety), the promoter for the granule bound starch synthase gene (GBSS) (Visser et al., Plant Mol. Biol. 17:691-699 (1991), herein incorporated by reference in its entirety) and other class I and II patatins promoters (Koster-Topfer et al., Mol Gen Genet. 219:390-396 (1989); Mignery et al., Gene. 62:27-44 (1988), both of which are herein incorporated by reference in their entirety).

Other promoters can also be used to express a methionine pathway protein or fragment thereof in specific tissues, such as seeds or fruits. The promoter for β-conglycinin (Chen et al., Dev. Genet. 10: 112-122 (1989), herein incorporated by reference in its entirety) or other seed-specific promoters such as the napin and phaseolin promoters, can be used. The zeins are a group of storage proteins found in maize endosperm. Genomic clones for zein genes have been isolated (Pedersen et al., Cell 29:1015-1026 (1982), herein incorporated by reference in its entirety) and the promoters from these clones, including the 15 kD, 16 kD, 19 kD, 22 kD, 27 kD and γ genes, could also be used. Other promoters known to function, for example, in maize include the promoters for the following genes: waxy, Brittle, Shrunken 2, Branching enzymes I and II, starch synthases, debranching enzymes, oleosins, glutelins and sucrose synthases. A particularly preferred promoter for maize endosperm expression is the promoter for the glutelin gene from rice, more particularly the Osgt-1 promoter (Zheng et al., Mol. Cell. Biol. 13:5829-5842 (1993), herein incorporated by reference in its entirety). Examples of promoters suitable for expression in wheat include those promoters for the ADPglucose pyrosynthase (ADPGPP) subunits, the granule bound and other starch synthase, the branching and debranching enzymes, the embryogenesis-abundant proteins, the gliadins and the glutenins. Examples of such promoters in rice include those promoters for the ADPGPP subunits, the granule bound and other starch synthase, the branching enzymes, the debranching enzymes, sucrose synthases and the glutelins. A particularly preferred promoter is the promoter for rice glutelin, Osgt-1. Examples of such promoters for barley include those for the ADPGPP subunits, the granule bound and other starch synthase, the branching enzymes, the debranching enzymes, sucrose synthases, the hordeins, the embryo globulins and the aleurone specific proteins.

Root specific promoters may also be used. An example of such a promoter is the promoter for the acid chitinase gene (Samac et al., Plant Mol. Biol. 25:587-596 (1994), the entirety of which is herein incorporated by reference). Expression in root tissue could also be accomplished by utilizing the root specific subdomains of the CaMV35S promoter that have been identified (Lam et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:7890-7894 (1989), herein incorporated by reference in its entirety). Other root cell specific promoters include those reported by Conkling et al. (Conkling et al., Plant Physiol. 93:1203-1211 (1990), the entirety of which is herein incorporated by reference).

Additional promoters that may be utilized are described, for example, in U.S. Pat. Nos. 5,378,619; 5,391,725; 5,428,147; 5,447,858; 5,608,144; 5,608,144; 5,614,399; 5,633,441; 5,633,435; and 4,633,436, all of which are herein incorporated in their entirety. In addition, a tissue specific enhancer may be used (Fromm et al., The Plant Cell 1:977-984 (1989), the entirety of which is herein incorporated by reference).

Constructs or vectors may also include with the coding region of interest a nucleic acid sequence that acts, in whole or in part, to terminate transcription of that region. For example, such sequences have been isolated including the Tr7 3′ sequence and the NOS 3′ sequence (Ingelbrecht et al., The Plant Cell 1:671-680 (1989), the entirety of which is herein incorporated by reference; Bevan et al., Nucleic Acids Res. 11:369-385 (1983), the entirety of which is herein incorporated by reference), or the like.

A vector or construct may also include regulatory elements. Examples of such include the Adh intron 1 (Callis et al., Genes and Develop. 1:1183-1200 (1987), the entirety of which is herein incorporated by reference), the sucrose synthase intron (Vasil et al., Plant Physiol. 91:1575-1579 (1989), the entirety of which is herein incorporated by reference) and the TMV omega element (Gallie et al., The Plant Cell 1:301-311 (1989), the entirety of which is herein incorporated by reference). These and other regulatory elements may be included when appropriate.

A vector or construct may also include a selectable marker. Selectable markers may also be used to select for plants or plant cells that contain the exogenous genetic material. Examples of such include, but are not limited to, a neo gene (Potrykus et al., Mol. Gen. Genet. 199:183-188 (1985), the entirety of which is herein incorporated by reference) which codes for kanamycin resistance and can be selected for using kanamycin, G418, etc.; a bar gene which codes for bialaphos resistance; a mutant EPSP synthase gene (Hinchee et al., Bio/Technology 6:915-922 (1988), the entirety of which is herein incorporated by reference) which encodes glyphosate resistance; a nitrilase gene which confers resistance to bromoxynil (Stalker et al., J. Biol. Chem. 263:6310-6314 (1988), the entirety of which is herein incorporated by reference); a mutant acetolactate synthase gene (ALS) which confers imidazolinone or sulphonylurea resistance (European Patent Application 154,204 (Sep. 11, 1985), the entirety of which is herein incorporated by reference); and a methotrexate resistant DHFR gene (Thillet et al., J. Biol. Chem. 263:12500-12508 (1988), the entirety of which is herein incorporated by reference).

A vector or construct may also include a transit peptide. Incorporation of a suitable chloroplast transit peptide may also be employed (European Patent Application Publication Number 0218571, the entirety of which is herein incorporated by reference). Translational enhancers may also be incorporated as part of the vector DNA. DNA constructs could contain one or more 5′ non-translated leader sequences which may serve to enhance expression of the gene products from the resulting mRNA transcripts. Such sequences may be derived from the promoter selected to express the gene or can be specifically modified to increase translation of the mRNA. Such regions may also be obtained from viral RNAs, from suitable eukaryotic genes, or from a synthetic gene sequence. For a review of optimizing expression of transgenes, see Koziel et al., Plant Mol. Biol. 32:393-405 (1996), the entirety of which is herein incorporated by reference.

A vector or construct may also include a screenable marker. Screenable markers may be used to monitor expression. Exemplary screenable markers include a β-glucuronidase or uidA gene (GUS) which encodes an enzyme for which various chromogenic substrates are known (Jefferson, Plant Mol. Biol, Rep. 5:387-405 (1987), the entirety of which is herein incorporated by reference; Jefferson et al., EMBO J. 6:3901-3907 (1987), the entirety of which is herein incorporated by reference); an R-locus gene, which encodes a product that regulates the production of anthocyanin pigments (red color) in plant tissues (Dellaporta et al., Stadler Symposium 11:263-282 (1988), the entirety of which is herein incorporated by reference); a lactamase gene (Sutcliffe et al., Proc. Natl. Acad. Sci. (U.S.A.) 75:3737-3741 (1978), the entirety of which is herein incorporated by reference), a gene which encodes an enzyme for which various chromogenic substrates are known (e.g., PADAC, a chromogenic cephalosporin); a luciferase gene (Ow et al., Science 234:856-859 (1986), the entirety of which is herein incorporated by reference); a xylE gene (Zukowsky et al., Proc. Natl. Acad. Sci. (U.S.A.) 80:1101-1105 (1983), the entirety of which is herein incorporated by reference) which encodes a catechol diozygenase that can convert chromogenic catechols; an α-amylase gene (Ikatu et al., Bio/Technol. 8:241-242 (1990), the entirety of which is herein incorporated by reference); a tyrosinase gene (Katz et al., J. Gen. Microbiol. 129:2703-2714 (1983), the entirety of which is herein incorporated by reference) which encodes an enzyme capable of oxidizing tyrosine to DOPA and dopaquinone which in turn condenses to melanin; an α-galactosidase, which will turn a chromogenic α-galactose substrate.

Included within the terms “selectable or screenable marker genes” are also genes which encode a secretable marker whose secretion can be detected as a means of identifying or selecting for transformed cells. Examples include markers which encode a secretable antigen that can be identified by antibody interaction, or even secretable enzymes which can be detected catalytically. Secretable proteins fall into a number of classes, including small, diffusible proteins which are detectable, (e.g., by ELISA), small active enzymes which are detectable in extracellular solution (e.g., α-amylase, β-lactamase, phosphinothricin transferase), or proteins which are inserted or trapped in the cell wall (such as proteins which include a leader sequence such as that found in the expression unit of extension or tobacco PR-S). Other possible selectable and/or screenable marker genes will be apparent to those of skill in the art.

There are many methods for introducing transforming nucleic acid molecules into plant cells. Suitable methods are believed to include virtually any method by which nucleic acid molecules may be introduced into a cell, such as by Agrobacterium infection or direct delivery of nucleic acid molecules such as, for example, by PEG-mediated transformation, by electroporation or by acceleration of DNA coated particles, etc (Potrykus, Ann. Rev. Plant Physiol. Plant Mol. Biol. 42:205-225 (1991), the entirety of which is herein incorporated by reference; Vasil, Plant Mol. Biol. 25:925-937 (1994), the entirety of which is herein incorporated by reference). For example, electroporation has been used to transform maize protoplasts (Fromm et al., Nature 312:791-793 (1986), the entirety of which is herein incorporated by reference).

Other vector systems suitable for introducing transforming DNA into a host plant cell include but are not limited to binary artificial chromosome (BIBAC) vectors (Hamilton et al., Gene 200:107-116 (1997), the entirety of which is herein incorporated by reference); and transfection with RNA viral vectors (Della-Cioppa et al., Ann. N.Y. Acad. Sci. (1996), 792 (Engineering Plants for Commercial Products and Applications), 57-61, the entirety of which is herein incorporated by reference). Additional vector systems also include plant selectable YAC vectors such as those described in Mullen et al., Molecular Breeding 4:449-457 (1988), the entirety of which is herein incorporated by reference).

Technology for introduction of DNA into cells is well known to those of skill in the art. Four general methods for delivering a gene into cells have been described: (1) chemical methods (Graham and van der Eb, Virology 54:536-539 (1973), the entirety of which is herein incorporated by reference); (2) physical methods such as microinjection (Capecchi, Cell 22:479-488 (1980), the entirety of which is herein incorporated by reference), electroporation (Wong and Neumann, Biochem. Biophys. Res. Commun. 107:584-587 (1982); Fromm et al., Proc. Natl. Acad. Sci. (U.S.A.) 82:5824-5828 (1985); U.S. Pat. No. 5,384,253, all of which are herein incorporated in their entirety); and the gene gun (Johnston and Tang, Methods Cell Biol. 43:353-365 (1994), the entirety of which is herein incorporated by reference); (3) viral vectors (Clapp, Clin. Perinatol. 20:155-168 (1993); Lu et al., J. Exp. Med. 178:2089-2096 (1993); Eglitis and Anderson, Biotechniques 6:608-614 (1988), all of which are herein incorporated in their entirety); and (4) receptor-mediated mechanisms (Curiel et al., Hum. Gen. Ther. 3:147-154 (1992), Wagner et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:6099-6103 (1992), both of which are incorporated by reference in their entirety).

Acceleration methods that may be used include, for example, microprojectile bombardment and the like. One example of a method for delivering transforming nucleic acid molecules to plant cells is microprojectile bombardment. This method has been reviewed by Yang and Christou (eds.), Particle Bombardment Technology for Gene Transfer, Oxford Press, Oxford, England (1994), the entirety of which is herein incorporated by reference). Non-biological particles (microprojectiles) that may be coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, gold, platinum and the like.

A particular advantage of microprojectile bombardment, in addition to it being an effective means of reproducibly transforming monocots, is that neither the isolation of protoplasts (Cristou et al., Plant Physiol. 87:671-674 (1988), the entirety of which is herein incorporated by reference) nor the susceptibility of Agrobacterium infection are required. An illustrative embodiment of a method for delivering DNA into maize cells by acceleration is a biolistics α-particle delivery system, which can be used to propel particles coated with DNA through a screen, such as a stainless steel or Nytex screen, onto a filter surface covered with corn cells cultured in suspension. Gordon-Kamm et al., describes the basic procedure for coating tungsten particles with DNA (Gordon-Kamm et al., Plant Cell 2:603-618 (1990), the entirety of which is herein incorporated by reference). The screen disperses the tungsten nucleic acid particles so that they are not delivered to the recipient cells in large aggregates. A particle delivery system suitable for use with the present invention is the helium acceleration PDS-1000/He gun is available from Bio-Rad Laboratories (Bio-Rad, Hercules, Calif.) (Sanford et al., Technique 3:3-16 (1991), the entirety of which is herein incorporated by reference).

For the bombardment, cells in suspension may be concentrated on filters. Filters containing the cells to be bombarded are positioned at an appropriate distance below the microprojectile stopping plate. If desired, one or more screens are also positioned between the gun and the cells to be bombarded.

Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the microprojectile stopping plate. If desired, one or more screens are also positioned between the acceleration device and the cells to be bombarded. Through the use of techniques set forth herein one may obtain up to 1000 or more foci of cells transiently expressing a marker gene. The number of cells in a focus which express the exogenous gene product 48 hours post-bombardment often range from one to ten and average one to three.

In bombardment transformation, one may optimize the pre-bombardment culturing conditions and the bombardment parameters to yield the maximum numbers of stable transformants. Both the physical and biological parameters for bombardment are important in this technology. Physical factors are those that involve manipulating the DNA/microprojectile precipitate or those that affect the flight and velocity of either the macro- or microprojectiles. Biological factors include all steps involved in manipulation of cells before and immediately after bombardment, the osmotic adjustment of target cells to help alleviate the trauma associated with bombardment and also the nature of the transforming DNA, such as linearized DNA or intact supercoiled plasmids. It is believed that pre-bombardment manipulations are especially important for successful transformation of immature embryos.

In another alternative embodiment, plastids can be stably transformed. Methods disclosed for plastid transformation in higher plants include the particle gun delivery of DNA containing a selectable marker and targeting of the DNA to the plastid genome through homologous recombination (Svab et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:8526-8530 (1990); Svab and Maliga, Proc. Natl. Acad. Sci. (U.S.A.) 90:913-917 (1993); Staub and Maliga, EMBO J. 12:601-606 (1993); U.S. Pat. Nos. 5,451,513 and 5,545,818, all of which are herein incorporated by reference in their entirety).

Accordingly, it is contemplated that one may wish to adjust various aspects of the bombardment parameters in small scale studies to fully optimize the conditions. One may particularly wish to adjust physical parameters such as gap distance, flight distance, tissue distance and helium pressure. One may also minimize the trauma reduction factors by modifying conditions which influence the physiological state of the recipient cells and which may therefore influence transformation and integration efficiencies. For example, the osmotic state, tissue hydration and the subculture stage or cell cycle of the recipient cells may be adjusted for optimum transformation. The execution of other routine adjustments will be known to those of skill in the art in light of the present disclosure.

Agrobacterium-mediated transfer is a widely applicable system for introducing genes into plant cells because the DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art. See, for example the methods described by Fraley et al., Bio/Technology 3:629-635 (1985) and Rogers et al., Methods Enzymol. 153:253-277 (1987), both of which are herein incorporated by reference in their entirety. Further, the integration of the Ti-DNA is a relatively precise process resulting in few rearrangements. The region of DNA to be transferred is defined by the border sequences and intervening DNA is usually inserted into the plant genome as described (Spielmann et al., Mol. Gen. Genet. 205:34 (1986), the entirety of which is herein incorporated by reference).

Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations as described (Klee et al., In: Plant DNA Infectious Agents, Hohn and Schell (eds.), Springer-Verlag, New York, pp. 179-203 (1985), the entirety of which is herein incorporated by reference. Moreover, technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes and are suitable for present purposes (Rogers et al., Methods Enzymol. 153:253-277 (1987)). In addition, Agrobacterium containing both armed and disarmed Ti genes can be used for the transformations. In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene transfer.

A transgenic plant formed using Agrobacterium transformation methods typically contains a single gene on one chromosome. Such transgenic plants can be referred to as being heterozygous for the added gene. More preferred is a transgenic plant that is homozygous for the added structural gene; i.e., a transgenic plant that contains two added genes, one gene at the same locus on each chromosome of a chromosome pair. A homozygous transgenic plant can be obtained by sexually mating (selfing) an independent segregant transgenic plant that contains a single added gene, germinating some of the seed produced and analyzing the resulting plants produced for the gene of interest.

It is also to be understood that two different transgenic plants can also be mated to produce offspring that contain two independently segregating added, exogenous genes. Selfing of appropriate progeny can produce plants that are homozygous for both added, exogenous genes that encode a polypeptide of interest. Back-crossing to a parental plant and out-crossing with a non-transgenic plant are also contemplated, as is vegetative propagation.

Transformation of plant protoplasts can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation and combinations of these treatments (See, for example, Potrykus et al., Mol. Gen. Genet. 205:193-200 (1986); Lorz et al., Mol. Gen. Genet. 199:178 (1985); Fromm et al., Nature 319:791 (1986); Uchimiya et al., Mol. Gen. Genet. 204:204 (1986); Marcotte et al., Nature 335:454-457 (1988), all of which are herein incorporated by reference in their entirety).

Application of these systems to different plant strains depends upon the ability to regenerate that particular plant strain from protoplasts. Illustrative methods for the regeneration of cereals from protoplasts are described (Fujimura et al., Plant Tissue Culture Letters 2:74 (1985); Toriyama et al., Theor Appl. Genet. 205:34 (1986); Yamada et al., Plant Cell Rep. 4:85 (1986); Abdullah et al., Biotechnolog 4:1087 (1986), all of which are herein incorporated by reference in their entirety).

To transform plant strains that cannot be successfully regenerated from protoplasts, other ways to introduce DNA into intact cells or tissues can be utilized. For example, regeneration of cereals from immature embryos or explants can be effected as described (Vasil, Biotechnology 6:397 (1988), the entirety of which is herein incorporated by reference). In addition, “particle gun” or high-velocity microprojectile technology can be utilized (Vasil et al., Bio/Technology 10:667 (1992), the entirety of which is herein incorporated by reference).

Using the latter technology, DNA is carried through the cell wall and into the cytoplasm on the surface of small metal particles as described (Klein et al., Nature 328:70 (1987); Klein et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:8502-8505 (1988); McCabe et al., Bio/Technology 6:923 (1988), all of which are herein incorporated by reference in their entirety). The metal particles penetrate through several layers of cells and thus allow the transformation of cells within tissue explants.

Other methods of cell transformation can also be used and include but are not limited to introduction of DNA into plants by direct DNA transfer into pollen (Zhou et al., Methods Enzymol. 101:433 (1983); Hess et al., Intern Rev. Cytol. 107:367 (1987); Luo et al., Plant Mol. Biol. Reporter 6:165 (1988), all of which are herein incorporated by reference in their entirety), by direct injection of DNA into reproductive organs of a plant (Pena et al., Nature 325:274 (1987), the entirety of which is herein incorporated by reference), or by direct injection of DNA into the cells of immature embryos followed by the rehydration of desiccated embryos (Neuhaus et al., Theor. Appl. Genet. 75:30 (1987), the entirety of which is herein incorporated by reference).

The regeneration, development and cultivation of plants from single plant protoplast transformants or from various transformed explants is well known in the art (Weissbach and Weissbach, In: Methods for Plant Molecular Biology, Academic Press, San Diego, Calif., (1988), the entirety of which is herein incorporated by reference). This regeneration and growth process typically includes the steps of selection of transformed cells, culturing those individualized cells through the usual stages of embryonic development through the rooted plantlet stage. Transgenic embryos and seeds are similarly regenerated. The resulting transgenic rooted shoots are thereafter planted in an appropriate plant growth medium such as soil.

The development or regeneration of plants containing the foreign, exogenous gene that encodes a protein of interest is well known in the art. Preferably, the regenerated plants are self-pollinated to provide homozygous transgenic plants. Otherwise, pollen obtained from the regenerated plants is crossed to seed-grown plants of agronomically important lines. Conversely, pollen from plants of these important lines is used to pollinate regenerated plants. A transgenic plant of the present invention containing a desired polypeptide is cultivated using methods well known to one skilled in the art.

There are a variety of methods for the regeneration of plants from plant tissue. The particular method of regeneration will depend on the starting plant tissue and the particular plant species to be regenerated.

Methods for transforming dicots, primarily by use of Agrobacterium tumefaciens and obtaining transgenic plants have been published for cotton (U.S. Pat. No. 5,004,863; U.S. Pat. No. 5,159,135; U.S. Pat. No. 5,518,908, all of which are herein incorporated by reference in their entirety); soybean (U.S. Pat. No. 5,569,834; U.S. Pat. No. 5,416,011; McCabe et. al., Biotechnology 6:923 (1988); Christou et al., Plant Physiol. 87:671-674 (1988); all of which are herein incorporated by reference in their entirety); Brassica (U.S. Pat. No. 5,463,174, the entirety of which is herein incorporated by reference); peanut (Cheng et al., Plant Cell Rep. 15:653-657 (1996), McKently et al., Plant Cell Rep. 14:699-703 (1995), all of which are herein incorporated by reference in their entirety); papaya; and pea (Grant et al., Plant Cell Rep. 15:254-258 (1995), the entirety of which is herein incorporated by reference).

Transformation of monocotyledons using electroporation, particle bombardment and Agrobacterium have also been reported. Transformation and plant regeneration have been achieved in asparagus (Bytebier et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:5354 (1987), the entirety of which is herein incorporated by reference); barley (Wan and Lemaux, Plant Physiol 104:37 (1994), the entirety of which is herein incorporated by reference); maize (Rhodes et al., Science 240:204 (1988); Gordon-Kamm et al., Plant Cell 2:603-618 (1990); Fromm et al., Bio/Technology 8:833 (1990); Koziel et al., Bio/Technology 11:194 (1993); Armstrong et al., Crop Science 35:550-557 (1995); all of which are herein incorporated by reference in their entirety); oat (Somers et al., Bio/Technology 10:1589 (1992), the entirety of which is herein incorporated by reference); orchard grass (Horn et al., Plant Cell Rep. 7:469 (1988), the entirety of which is herein incorporated by reference); rice (Toriyama et al., Theor Appl. Genet. 205:34 (1986); Part et al., Plant Mol. Biol. 32:1135-1148 (1996); Abedinia et al., Aust. J. Plant Physiol. 24:133-141 (1997); Zhang and Wu, Theor. Appl. Genet. 76:835 (1988); Zhang et al., Plant Cell Rep. 7:379 (1988); Battraw and Hall, Plant Sci. 86:191-202 (1992); Christou et al., Bio/Technology 9:957 (1991), all of which are herein incorporated by reference in their entirety); rye (De la Pena et al., Nature 325:274 (1987), the entirety of which is herein incorporated by reference); sugarcane (Bower and Birch, Plant J. 2:409 (1992), the entirety of which is herein incorporated by reference); tall fescue (Wang et al., Bio/Technology 10:691 (1992), the entirety of which is herein incorporated by reference) and wheat (Vasil et al., Bio/Technology 10:667 (1992), the entirety of which is herein incorporated by reference; U.S. Pat. No. 5,631,152, the entirety of which is herein incorporated by reference.)

Assays for gene expression based on the transient expression of cloned nucleic acid constructs have been developed by introducing the nucleic acid molecules into plant cells by polyethylene glycol treatment, electroporation, or particle bombardment (Marcotte et al., Nature 335:454-457 (1988), the entirety of which is herein incorporated by reference; Marcotte et al., Plant Cell 1:523-532 (1989), the entirety of which is herein incorporated by reference; McCarty et al., Cell 66:895-905 (1991), the entirety of which is herein incorporated by reference; Hattori et al., Genes Dev. 6:609-618 (1992), the entirety of which is herein incorporated by reference; Goff et al., EMBO J. 9:2517-2522 (1990), the entirety of which is herein incorporated by reference). Transient expression systems may be used to functionally dissect gene constructs (see generally, Mailga et al., Methods in Plant Molecular Biology, Cold Spring Harbor Press (1995)).

Any of the nucleic acid molecules of the present invention may be introduced into a plant cell in a permanent or transient manner in combination with other genetic elements such as vectors, promoters, enhancers etc. Further, any of the nucleic acid molecules of the present invention may be introduced into a plant cell in a manner that allows for overexpression of the protein or fragment thereof encoded by the nucleic acid molecule.

Cosuppression is the reduction in expression levels, usually at the level of RNA, of a particular endogenous gene or gene family by the expression of a homologous sense construct that is capable of transcribing mRNA of the same strandedness as the transcript of the endogenous gene (Napoli et al., Plant Cell 2:279-289 (1990), the entirety of which is herein incorporated by reference; van der Krol et al., Plant Cell 2:291-299 (1990), the entirety of which is herein incorporated by reference). Cosuppression may result from stable transformation with a single copy nucleic acid molecule that is homologous to a nucleic acid sequence found with the cell (Prolls and Meyer, Plant J. 2:465-475 (1992), the entirety of which is herein incorporated by reference) or with multiple copies of a nucleic acid molecule that is homologous to a nucleic acid sequence found with the cell (Mittlesten et al., Mol. Gen. Genet. 244:325-330 (1994), the entirety of which is herein incorporated by reference). Genes, even though different, linked to homologous promoters may result in the cosuppression of the linked genes (Vaucheret, C. R. Acad. Sci. III 316:1471-1483 (1993), the entirety of which is herein incorporated by reference).

This technique has, for example, been applied to generate white flowers from red petunia and tomatoes that do not ripen on the vine. Up to 50% of petunia transformants that contained a sense copy of the glucoamylase (CHS) gene produced white flowers or floral sectors; this was as a result of the post-transcriptional loss of mRNA encoding CHS (Flavell, Proc. Natl. Acad. Sci. (U.S.A.) 91:3490-3496 (1994), the entirety of which is herein incorporated by reference); van Blokland et al., Plant J. 6:861-877 (1994), the entirety of which is herein incorporated by reference). Cosuppression may require the coordinate transcription of the transgene and the endogenous gene and can be reset by a developmental control mechanism (Jorgensen, Trends Biotechnol. 8:340-344 (1990), the entirety of which is herein incorporated by reference; Meins and Kunz, In: Gene Inactivation and Homologous Recombination in Plants, Paszkowski (ed.), pp. 335-348, Kluwer Academic, Netherlands (1994), the entirety of which is herein incorporated by reference).

It is understood that one or more of the nucleic acids of the present invention may be introduced into a plant cell and transcribed using an appropriate promoter with such transcription resulting in the cosuppression of an endogenous methionine pathway protein.

Antisense approaches are a way of preventing or reducing gene function by targeting the genetic material (Mol et al., FEBS Lett. 268:427-430 (1990), the entirety of which is herein incorporated by reference). The objective of the antisense approach is to use a sequence complementary to the target gene to block its expression and create a mutant cell line or organism in which the level of a single chosen protein is selectively reduced or abolished. Antisense techniques have several advantages over other ‘reverse genetic’ approaches. The site of inactivation and its developmental effect can be manipulated by the choice of promoter for antisense genes or by the timing of external application or microinjection. Antisense can manipulate its specificity by selecting either unique regions of the target gene or regions where it shares homology to other related genes (Hiatt et al., In: Genetic Engineering, Setlow (ed.), Vol. 11, New York: Plenum 49-63 (1989), the entirety of which is herein incorporated by reference).

The principle of regulation by antisense RNA is that RNA that is complementary to the target mRNA is introduced into cells, resulting in specific RNA:RNA duplexes being formed by base pairing between the antisense substrate and the target mRNA (Green et al., Annu. Rev. Biochem. 55:569-597 (1986), the entirety of which is herein incorporated by reference). Under one embodiment, the process involves the introduction and expression of an antisense gene sequence. Such a sequence is one in which part or all of the normal gene sequences are placed under a promoter in inverted orientation so that the ‘wrong’ or complementary strand is transcribed into a noncoding antisense RNA that hybridizes with the target mRNA and interferes with its expression (Takayama and Inouye, Crit. Rev. Biochem. Mol. Biol. 25:155-184 (1990), the entirety of which is herein incorporated by reference). An antisense vector is constructed by standard procedures and introduced into cells by transformation, transfection, electroporation, microinjection, infection, etc. The type of transformation and choice of vector will determine whether expression is transient or stable. The promoter used for the antisense gene may influence the level, timing, tissue, specificity, or inducibility of the antisense inhibition.

It is understood that the activity of a methionine pathway protein in a plant cell may be reduced or depressed by growing a transformed plant cell containing a nucleic acid molecule whose non-transcribed strand encodes a methionine pathway protein or fragment thereof.

Antibodies have been expressed in plants (Hiatt et al., Nature 342:76-78 (1989), the entirety of which is herein incorporated by reference; Conrad and Fielder, Plant Mol. Biol. 26:1023-1030 (1994), the entirety of which is herein incorporated by reference). Cytoplamsic expression of a scFv (single-chain Fv antibodies) has been reported to delay infection by artichoke mottled crinkle virus. Transgenic plants that express antibodies directed against endogenous proteins may exhibit a physiological effect (Philips et al., EMBO J. 16:4489-4496 (1997), the entirety of which is herein incorporated by reference; Marion-Poll, Trends in Plant Science 2:447-448 (1997), the entirety of which is herein incorporated by reference). For example, expressed anti-abscisic antibodies have been reported to result in a general perturbation of seed development (Philips et al., EMBO J. 16: 4489-4496 (1997)).

Antibodies that are catalytic may also be expressed in plants (abzymes). The principle behind abzymes is that since antibodies may be raised against many molecules, this recognition ability can be directed toward generating antibodies that bind transition states to force a chemical reaction forward (Persidas, Nature Biotechnology 15:1313-1315 (1997), the entirety of which is herein incorporated by reference; Baca et al., Ann. Rev. Biophys. Biomol. Struct. 26:461-493 (1997), the entirety of which is herein incorporated by reference). The catalytic abilities of abzymes may be enhanced by site directed mutagenesis. Examples of abzymes are, for example, set forth in U.S. Pat. No. 5,658,753; U.S. Pat. No. 5,632,990; U.S. Pat. No. 5,631,137; U.S. Pat. No. 5,602,015; U.S. Pat. No. 5,559,538; U.S. Pat. No. 5,576,174; U.S. Pat. No. 5,500,358; U.S. Pat. No. 5,318,897; U.S. Pat. No. 5,298,409; U.S. Pat. No. 5,258,289 and U.S. Pat. No. 5,194,585, all of which are herein incorporated in their entirety.

It is understood that any of the antibodies of the present invention may be expressed in plants and that such expression can result in a physiological effect. It is also understood that any of the expressed antibodies may be catalytic.

(b) Fungal Constructs and Fungal Transformants

The present invention also relates to a fungal recombinant vector comprising exogenous genetic material. The present invention also relates to a fungal cell comprising a fungal recombinant vector. The present invention also relates to methods for obtaining a recombinant fungal host cell comprising introducing into a fungal host cell exogenous genetic material.

Exogenous genetic material may be transferred into a fungal cell. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof or fragments of either. The fungal recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures. The choice of a vector will typically depend on the compatibility of the vector with the fungal host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the fungal host.

The fungal vector may be an autonomously replicating vector, i.e., a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any means for assuring self-replication. Alternatively, the vector may be one which, when introduced into the fungal cell, is integrated into the genome and replicated together with the chromosome(s) into which it has been integrated. For integration, the vector may rely on the nucleic acid sequence of the vector for stable integration of the vector into the genome by homologous or nonhomologous recombination. Alternatively, the vector may contain additional nucleic acid sequences for directing integration by homologous recombination into the genome of the fungal host. The additional nucleic acid sequences enable the vector to be integrated into the host cell genome at a precise location(s) in the chromosome(s). To increase the likelihood of integration at a precise location, there should be preferably two nucleic acid sequences which individually contain a sufficient number of nucleic acids, preferably 400 bp to 1500 bp, more preferably 800 bp to 1000 bp, which are highly homologous with the corresponding target sequence to enhance the probability of homologous recombination. These nucleic acid sequences may be any sequence that is homologous with a target sequence in the genome of the fungal host cell and, furthermore, may be non-encoding or encoding sequences.

For autonomous replication, the vector may further comprise an origin of replication enabling the vector to replicate autonomously in the host cell in question. Examples of origin of replications for use in a yeast host cell are the 2 micron origin of replication and the combination of CEN3 and ARS 1. Any origin of replication may be used which is compatible with the fungal host cell of choice.

The fungal vectors of the present invention preferably contain one or more selectable markers which permit easy selection of transformed cells. A selectable marker is a gene the product of which provides, for example biocide or viral resistance, resistance to heavy metals, prototrophy to auxotrophs and the like. The selectable marker may be selected from the group including, but not limited to, amdS (acetamidase), argB (ornithine carbamoyltransferase), bar (phosphinothricin acetyltransferase), hygB (hygromycin phosphotransferase), niaD (nitrate reductase), pyrG (orotidine-5′-phosphate decarboxylase) and sC (sulfate adenyltransferase) and trpC (anthranilate synthase). Preferred for use in an Aspergillus cell are the amdS and pyrG markers of Aspergillus nidulans or Aspergillus oryzae and the bar marker of Streptomyces hygroscopicus. Furthermore, selection may be accomplished by co-transformation, e.g., as described in WO 91/17243, the entirety of which is herein incorporated by reference. A nucleic acid sequence of the present invention may be operably linked to a suitable promoter sequence. The promoter sequence is a nucleic acid sequence which is recognized by the fungal host cell for expression of the nucleic acid sequence. The promoter sequence contains transcription and translation control sequences which mediate the expression of the protein or fragment thereof.

A promoter may be any nucleic acid sequence which shows transcriptional activity in the fungal host cell of choice and may be obtained from genes encoding polypeptides either homologous or heterologous to the host cell. Examples of suitable promoters for directing the transcription of a nucleic acid construct of the invention in a filamentous fungal host are promoters obtained from the genes encoding Aspergillus oryzae TAKA amylase, Rhizomucor miehei aspartic proteinase, Aspergillus niger neutral alpha-amylase, Aspergillus niger acid stable alpha-amylase, Aspergillus niger or Aspergillus awamori glucoamylase (glaA), Rhizomucor miehei lipase, Aspergillus oryzae alkaline protease, Aspergillus oryzae triose phosphate isomerase, Aspergillus nidulans acetamidase and hybrids thereof. In a yeast host, a useful promoter is the Saccharomyces cerevisiae enolase (eno-1) promoter. Particularly preferred promoters are the TAKA amylase, NA2-tpi (a hybrid of the promoters from the genes encoding Aspergillus niger neutral alpha-amylase and Aspergillus oryzae triose phosphate isomerase) and glaA promoters.

A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be operably linked to a terminator sequence at its 3′ terminus. The terminator sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any terminator which is functional in the fungal host cell of choice may be used in the present invention, but particularly preferred terminators are obtained from the genes encoding Aspergillus oryzae TAKA amylase, Aspergillus niger glucoamylase, Aspergillus nidulans anthranilate synthase, Aspergillus niger alpha-glucosidase and Saccharomyces cerevisiae enolase.

A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be operably linked to a suitable leader sequence. A leader sequence is a nontranslated region of a mRNA which is important for translation by the fungal host. The leader sequence is operably linked to the 5′ terminus of the nucleic acid sequence encoding the protein or fragment thereof. The leader sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any leader sequence which is functional in the fungal host cell of choice may be used in the present invention, but particularly preferred leaders are obtained from the genes encoding Aspergillus oryzae TAKA amylase and Aspergillus oryzae triose phosphate isomerase.

A polyadenylation sequence may also be operably linked to the 3′ terminus of the nucleic acid sequence of the present invention. The polyadenylation sequence is a sequence which when transcribed is recognized by the fungal host to add polyadenosine residues to transcribed mRNA. The polyadenylation sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any polyadenylation sequence which is functional in the fungal host of choice may be used in the present invention, but particularly preferred polyadenylation sequences are obtained from the genes encoding Aspergillus oryzae TAKA amylase, Aspergillus niger glucoamylase, Aspergillus nidulans anthranilate synthase and Aspergillus niger alpha-glucosidase.

To avoid the necessity of disrupting the cell to obtain the protein or fragment thereof and to minimize the amount of possible degradation of the expressed protein or fragment thereof within the cell, it is preferred that expression of the protein or fragment thereof gives rise to a product secreted outside the cell. To this end, a protein or fragment thereof of the present invention may be linked to a signal peptide linked to the amino terminus of the protein or fragment thereof. A signal peptide is an amino acid sequence which permits the secretion of the protein or fragment thereof from the fungal host into the culture medium. The signal peptide may be native to the protein or fragment thereof of the invention or may be obtained from foreign sources. The 5′ end of the coding sequence of the nucleic acid sequence of the present invention may inherently contain a signal peptide coding region naturally linked in translation reading frame with the segment of the coding region which encodes the secreted protein or fragment thereof. Alternatively, the 5′ end of the coding sequence may contain a signal peptide coding region which is foreign to that portion of the coding sequence which encodes the secreted protein or fragment thereof. The foreign signal peptide may be required where the coding sequence does not normally contain a signal peptide coding region. Alternatively, the foreign signal peptide may simply replace the natural signal peptide to obtain enhanced secretion of the desired protein or fragment thereof. The foreign signal peptide coding region may be obtained from a glucoamylase or an amylase gene from an Aspergillus species, a lipase or proteinase gene from Rhizomucor miehei, the gene for the alpha-factor from Saccharomyces cerevisiae, or the calf preprochymosin gene. An effective signal peptide for fungal host cells is the Aspergillus oryzae TAKA amylase signal, Aspergillus niger neutral amylase signal, the Rhizomucor miehei aspartic proteinase signal, the Humicola lanuginosus cellulase signal, or the Rhizomucor miehei lipase signal. However, any signal peptide capable of permitting secretion of the protein or fragment thereof in a fungal host of choice may be used in the present invention.

A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be linked to a propeptide coding region. A propeptide is an amino acid sequence found at the amino terminus of a proprotein or proenzyme. Cleavage of the propeptide from the proprotein yields a mature biochemically active protein. The resulting polypeptide is known as a propolypeptide or proenzyme (or a zymogen in some cases). Propolypeptides are generally inactive and can be converted to mature active polypeptides by catalytic or autocatalytic cleavage of the propeptide from the propolypeptide or proenzyme. The propeptide coding region may be native to the protein or fragment thereof or may be obtained from foreign sources. The foreign propeptide coding region may be obtained from the Saccharomyces cerevisiae alpha-factor gene or Myceliophthora thermophila laccase gene (WO 95/33836, the entirety of which is herein incorporated by reference).

The procedures used to ligate the elements described above to construct the recombinant expression vector of the present invention are well known to one skilled in the art (see, for example, Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd ed., Cold Spring Harbor, N.Y., (1989)).

The present invention also relates to recombinant fungal host cells produced by the methods of the present invention which are advantageously used with the recombinant vector of the present invention. The cell is preferably transformed with a vector comprising a nucleic acid sequence of the invention followed by integration of the vector into the host chromosome. The choice of fungal host cells will to a large extent depend upon the gene encoding the protein or fragment thereof and its source. The fungal host cell may, for example, be a yeast cell or a filamentous fungal cell.

“Yeast” as used herein includes Ascosporogenous yeast (Endomycetales), Basidiosporogenous yeast and yeast belonging to the Fungi Imperfecti (Blastomycetes). The Ascosporogenous yeasts are divided into the families Spermophthoraceae and Saccharomycetaceae. The latter is comprised of four subfamilies, Schizosaccharomycoideae (for example, genus Schizosaccharomyces), Nadsonioideae, Lipomycoideae and Saccharomycoideae (for example, genera Pichia, Kluyveromyces and Saccharomyces). The Basidiosporogenous yeasts include the genera Leucosporidim, Rhodosporidium, Sporidiobolus, Filobasidium and Filobasidiella. Yeast belonging to the Fungi Imperfecti are divided into two families, Sporobolomycetaceae (for example, genera Sorobolomyces and Bullera) and Cryptococcaceae (for example, genus Candida). Since the classification of yeast may change in the future, for the purposes of this invention, yeast shall be defined as described in Biology and Activities of Yeast (Skinner et al., Soc. App. Bacteriol. Symposium Series No. 9, (1980), the entirety of which is herein incorporated by reference). The biology of yeast and manipulation of yeast genetics are well known in the art (see, for example, Biochemistry and Genetics of Yeast, Bacil et al. (ed.), 2nd edition, 1987; The Yeasts, Rose and Harrison (eds.), 2nd ed., (1987); and The Molecular Biology of the Yeast Saccharomyces, Strathern et al. (eds.), (1981), all of which are herein incorporated by reference in their entirety).

“Fungi” as used herein includes the phyla Ascomycota, Basidiomycota, Chytridiomycota and Zygomycota (as defined by Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK; the entirety of which is herein incorporated by reference) as well as the Oomycota (as cited in Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK) and all mitosporic fungi (Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK). Representative groups of Ascomycota include, for example, Neurospora, Eupenicillium (=Penicillium), Emericella (=Aspergillus), Eurotiun (=Aspergillus) and the true yeasts listed above. Examples of Basidiomycota include mushrooms, rusts and smuts. Representative groups of Chytridiomycota include, for example, Allomyces, Blastocladiella, Coelomomyces and aquatic fungi. Representative groups of Oomycota include, for example, Saprolegniomycetous aquatic fungi (water molds) such as Achlya. Examples of mitosporic fungi include Aspergillus, Penicilliun, Candida and Alternaria. Representative groups of Zygomycota include, for example, Rhizopus and Mucor.

“Filamentous fungi” include all filamentous forms of the subdivision Eumycota and Oomycota (as defined by Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK). The filamentous fungi are characterized by a vegetative mycelium composed of chitin, cellulose, glucan, chitosan, mannan and other complex polysaccharides. Vegetative growth is by hyphal elongation and carbon catabolism is obligately aerobic. In contrast, vegetative growth by yeasts such as Saccharomyces cerevisiae is by budding of a unicellular thallus and carbon catabolism may be fermentative.

In one embodiment, the fungal host cell is a yeast cell. In a preferred embodiment, the yeast host cell is a cell of the species of Candida, Kluyveromyces, Saccharomyces, Schizosaccharomyces, Pichia and Yarrowia. In a preferred embodiment, the yeast host cell is a Saccharomyces cerevisiae cell, a Saccharomyces carlsbergensis, Saccharomyces diastaticus cell, a Saccharomyces douglasii cell, a Saccharomyces kluyveri cell, a Saccharomyces norbensis cell, or a Saccharomyces oviformis cell. In another preferred embodiment, the yeast host cell is a Kluyveromyces lactis cell. In another preferred embodiment, the yeast host cell is a Yarrowia lipolytica cell.

In another embodiment, the fungal host cell is a filamentous fungal cell. In a preferred embodiment, the filamentous fungal host cell is a cell of the species of, but not limited to, Acremonium, Aspergillus, Fusarium, Humicola, Myceliophthora, Mucor, Neurospora, Penicillium, Thielavia, Tolypocladium and Trichoderma. In a preferred embodiment, the filamentous fungal host cell is an Aspergillus cell. In another preferred embodiment, the filamentous fungal host cell is an Acremonium cell. In another preferred embodiment, the filamentous fungal host cell is a Fusarium cell. In another preferred embodiment, the filamentous fungal host cell is a Humicola cell. In another preferred embodiment, the filamentous fungal host cell is a Myceliophthora cell. In another even preferred embodiment, the filamentous fungal host cell is a Mucor cell. In another preferred embodiment, the filamentous fungal host cell is a Neurospora cell. In another preferred embodiment, the filamentous fungal host cell is a Penicillium cell. In another preferred embodiment, the filamentous fungal host cell is a Thielavia cell. In another preferred embodiment, the filamentous fungal host cell is a Tolypocladiun cell. In another preferred embodiment, the filamentous fungal host cell is a Trichoderma cell. In a preferred embodiment, the filamentous fungal host cell is an Aspergillus oryzae cell, an Aspergillus niger cell, an Aspergillus foetidus cell, or an Aspergillus japonicus cell. In another preferred embodiment, the filamentous fungal host cell is a Fusarium oxysporum cell or a Fusarium graminearum cell. In another preferred embodiment, the filamentous fungal host cell is a Humicola insolens cell or a Humicola lanuginosus cell. In another preferred embodiment, the filamentous fungal host cell is a Myceliophthora thermophila cell. In a most preferred embodiment, the filamentous fungal host cell is a Mucor miehei cell. In a most preferred embodiment, the filamentous fungal host cell is a Neurospora crassa cell. In a most preferred embodiment, the filamentous fungal host cell is a Penicillium purpurogenum cell. In another most preferred embodiment, the filamentous fungal host cell is a Thielavia terrestris cell. In another most preferred embodiment, the Trichoderma cell is a Trichoderma reesei cell, a Trichoderma viride cell, a Trichoderma longibrachiatum cell, a Trichoderma harzianum cell, or a Trichoderma koningii cell. In a preferred embodiment, the fungal host cell is selected from an A. nidulans cell, an A. niger cell, an A. oryzae cell and an A. sojae cell. In a further preferred embodiment, the fungal host cell is an A. nidulans cell.

The recombinant fungal host cells of the present invention may further comprise one or more sequences which encode one or more factors that are advantageous in the expression of the protein or fragment thereof, for example, an activator (e.g., a trans-acting factor), a chaperone and a processing protease. The nucleic acids encoding one or more of these factors are preferably not operably linked to the nucleic acid encoding the protein or fragment thereof. An activator is a protein which activates transcription of a nucleic acid sequence encoding a polypeptide (Kudla et al., EMBO 9:1355-1364 (1990); Jarai and Buxton, Current Genetics 26:2238-244 (1994); Verdier, Yeast 6:271-297 (1990), all of which are herein incorporated by reference in their entirety). The nucleic acid sequence encoding an activator may be obtained from the genes encoding Saccharomyces cerevisiae heme activator protein 1 (hap1), Saccharomyces cerevisiae galactose metabolizing protein 4 (gal4) and Aspergillus nidulans ammonia regulation protein (areA). For further examples, see Verdier, Yeast 6:271-297 (1990); MacKenzie et al., Journal of Gen. Microbiol. 139:2295-2307 (1993), both of which are herein incorporated by reference in their entirety). A chaperone is a protein which assists another protein in folding properly (Hartl et al., TIBS 19:20-25 (1994); Bergeron et al., TIBS 19:124-128 (1994); Demolder et al., J. Biotechnology 32:179-189 (1994); Craig, Science 260:1902-1903 (1993); Gething and Sambrook, Nature 355:33-45 (1992); Puig and Gilbert, J. Biol. Chem. 269:7764-7771 (1994); Wang and Tsou, FASEB Journal 7:1515-11157 (1993); Robinson et al., Bio/Technology 1:381-384 (1994), all of which are herein incorporated by reference in their entirety). The nucleic acid sequence encoding a chaperone may be obtained from the genes encoding Aspergillus oryzae protein disulphide isomerase, Saccharomyces cerevisiae calnexin, Saccharomyces cerevisiae BiP/GRP78 and Saccharomyces cerevisiae Hsp70. For further examples, see Gething and Sambrook, Nature 355:33-45 (1992); Hartl et al., TIBS 19:20-25 (1994). A processing protease is a protease that cleaves a propeptide to generate a mature biochemically active polypeptide (Enderlin and Ogrydziak, Yeast 10:67-79 (1994); Fuller et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:1434-1438 (1989); Julius et al., Cell 37:1075-1089 (1984); Julius et al., Cell 32:839-852 (1983), all of which are incorporated by reference in their entirety). The nucleic acid sequence encoding a processing protease may be obtained from the genes encoding Aspergillus niger Kex2, Saccharomyces cerevisiae dipeptidylaminopeptidase, Saccharomyces cerevisiae Kex2 and Yarrowia lipolytica dibasic processing endoprotease (xpr6). Any factor that is functional in the fungal host cell of choice may be used in the present invention.

Fungal cells may be transformed by a process involving protoplast formation, transformation of the protoplasts and regeneration of the cell wall in a manner known per se. Suitable procedures for transformation of Aspergillus host cells are described in EP 238 023 and Yelton et al., Proc. Natl. Acad. Sci. (U.S.A.) 81:1470-1474 (1984), both of which are herein incorporated by reference in their entirety. A suitable method of transforming Fusarium species is described by Malardier et al., Gene 78:147-156 (1989), the entirety of which is herein incorporated by reference. Yeast may be transformed using the procedures described by Becker and Guarente, In: Abelson and Simon, (eds.), Guide to Yeast Genetics and Molecular Biology, Methods Enzymol. Volume 194, pp 182-187, Academic Press, Inc., New York; Ito et al., J. Bacteriology 153:163 (1983); Hinnen et al., Proc. Natl. Acad. Sci. (U.S.A.) 75:1920 (1978), all of which are herein incorporated by reference in their entirety.

The present invention also relates to methods of producing the protein or fragment thereof comprising culturing the recombinant fungal host cells under conditions conducive for expression of the protein or fragment thereof. The fungal cells of the present invention are cultivated in a nutrient medium suitable for production of the protein or fragment thereof using methods known in the art. For example, the cell may be cultivated by shake flask cultivation, small-scale or large-scale fermentation (including continuous, batch, fed-batch, or solid state fermentations) in laboratory or industrial fermentors performed in a suitable medium and under conditions allowing the protein or fragment thereof to be expressed and/or isolated. The cultivation takes place in a suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using procedures known in the art (see, e.g., Bennett and LaSure (eds.), More Gene Manipulations in Fungi, Academic Press, CA, (1991), the entirety of which is herein incorporated by reference). Suitable media are available from commercial suppliers or may be prepared according to published compositions (e.g., in catalogues of the American Type Culture Collection, Manassas, Va.). If the protein or fragment thereof is secreted into the nutrient medium, a protein or fragment thereof can be recovered directly from the medium. If the protein or fragment thereof is not secreted, it is recovered from cell lysates.

The expressed protein or fragment thereof may be detected using methods known in the art that are specific for the particular protein or fragment. These detection methods may include the use of specific antibodies, formation of an enzyme product, or disappearance of an enzyme substrate. For example, if the protein or fragment thereof has enzymatic activity, an enzyme assay may be used. Alternatively, if polyclonal or monoclonal antibodies specific to the protein or fragment thereof are available, immunoassays may be employed using the antibodies to the protein or fragment thereof. The techniques of enzyme assay and immunoassay are well known to those skilled in the art.

The resulting protein or fragment thereof may be recovered by methods known in the arts. For example, the protein or fragment thereof may be recovered from the nutrient medium by conventional procedures including, but not limited to, centrifugation, filtration, extraction, spray-drying, evaporation, or precipitation. The recovered protein or fragment thereof may then be further purified by a variety of chromatographic procedures, e.g., ion exchange chromatography, gel filtration chromatography, affinity chromatography, or the like.

(c) Mammalian Constructs and Transformed Mammalian Cells

The present invention also relates to methods for obtaining a recombinant mammalian host cell, comprising introducing into a mammalian host cell exogenous genetic material. The present invention also relates to a mammalian cell comprising a mammalian recombinant vector. The present invention also relates to methods for obtaining a recombinant mammalian host cell, comprising introducing into a mammalian cell exogenous genetic material.

Mammalian cell lines available as hosts for expression are known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC, Manassas, Va.), such as HeLa cells, Chinese hamster ovary (CHO) cells, baby hamster kidney (BHK) cells and a number of other cell lines. Suitable promoters for mammalian cells are also known in the art and include viral promoters such as that from Simian Virus 40 (SV40) (Fiers et al., Nature 273:113 (1978), the entirety of which is herein incorporated by reference), Rous sarcoma virus (RSV), adenovirus (ADV) and bovine papilloma virus (BPV). Mammalian cells may also require terminator sequences and poly-A addition sequences. Enhancer sequences which increase expression may also be included and sequences which promote amplification of the gene may also be desirable (for example methotrexate resistance genes).

Vectors suitable for replication in mammalian cells may include viral replicons, or sequences which insure integration of the appropriate sequences encoding HCV epitopes into the host genome. For example, another vector used to express foreign DNA is vaccinia virus. In this case, for example, a nucleic acid molecule encoding a protein or fragment thereof is inserted into the vaccinia genome. Techniques for the insertion of foreign DNA into the vaccinia virus genome are known in the art and may utilize, for example, homologous recombination. Such heterologous DNA is generally inserted into a gene which is non-essential to the virus, for example, the thymidine kinase gene (tk), which also provides a selectable marker. Plasmid vectors that greatly facilitate the construction of recombinant viruses have been described (see, for example, Mackett et al., J Virol. 49:857 (1984); Chakrabarti et al., Mol. Cell. Biol. 5:3403 (1985); Moss, In: Gene Transfer Vectors For Mammalian Cells (Miller and Calos, eds., Cold Spring Harbor Laboratory, N.Y., p. 10, (1987); all of which are herein incorporated by reference in their entirety). Expression of the HCV polypeptide then occurs in cells or animals which are infected with the live recombinant vaccinia virus.

The sequence to be integrated into the mammalian sequence may be introduced into the primary host by any convenient means, which includes calcium precipitated DNA, spheroplast fusion, transformation, electroporation, biolistics, lipofection, microinjection, or other convenient means. Where an amplifiable gene is being employed, the amplifiable gene may serve as the selection marker for selecting hosts into which the amplifiable gene has been introduced. Alternatively, one may include with the amplifiable gene another marker, such as a drug resistance marker, e.g. neomycin resistance (G418 in mammalian cells), hygromycin in resistance etc., or an auxotrophy marker (HIS3, TRP1, LEU2, URA3, ADE2, LYS2, etc.) for use in yeast cells.

Depending upon the nature of the modification and associated targeting construct, various techniques may be employed for identifying targeted integration. Conveniently, the DNA may be digested with one or more restriction enzymes and the fragments probed with an appropriate DNA fragment which will identify the properly sized restriction fragment associated with integration.

One may use different promoter sequences, enhancer sequences, or other sequence which will allow for enhanced levels of expression in the expression host. Thus, one may combine an enhancer from one source, a promoter region from another source, a 5′-noncoding region upstream from the initiation methionine from the same or different source as the other sequences and the like. One may provide for an intron in the non-coding region with appropriate splice sites or for an alternative 3′-untranslated sequence or polyadenylation site. Depending upon the particular purpose of the modification, any of these sequences may be introduced, as desired.

Where selection is intended, the sequence to be integrated will have with it a marker gene, which allows for selection. The marker gene may conveniently be downstream from the target gene and may include resistance to a cytotoxic agent, e.g. antibiotics, heavy metals, or the like, resistance or susceptibility to HAT, gancyclovir, etc., complementation to an auxotrophic host, particularly by using an auxotrophic yeast as the host for the subject manipulations, or the like. The marker gene may also be on a separate DNA molecule, particularly with primary mammalian cells. Alternatively, one may screen the various transformants, due to the high efficiency of recombination in yeast, by using hybridization analysis, PCR, sequencing, or the like.

For homologous recombination, constructs can be prepared where the amplifiable gene will be flanked, normally on both sides with DNA homologous with the DNA of the target region. Depending upon the nature of the integrating DNA and the purpose of the integration, the homologous DNA will generally be within 100 kb, usually 50 kb, preferably about 25 kb, of the transcribed region of the target gene, more preferably within 2 kb of the target gene. Where modeling of the gene is intended, homology will usually be present proximal to the site of the mutation. The homologous DNA may include the 5′-upstream region outside of the transcriptional regulatory region or comprising any enhancer sequences, transcriptional initiation sequences, adjacent sequences, or the like. The homologous region may include a portion of the coding region, where the coding region may be comprised only of an open reading frame or combination of exons and introns. The homologous region may comprise all or a portion of an intron, where all or a portion of one or more exons may also be present. Alternatively, the homologous region may comprise the 3′-region, so as to comprise all or a portion of the transcriptional termination region, or the region 3′ of this region. The homologous regions may extend over all or a portion of the target gene or be outside the target gene comprising all or a portion of the transcriptional regulatory regions and/or the structural gene.

The integrating constructs may be prepared in accordance with conventional ways, where sequences may be synthesized, isolated from natural sources, manipulated, cloned, ligated, subjected to in vitro mutagenesis, primer repair, or the like. At various stages, the joined sequences may be cloned and analyzed by restriction analysis, sequencing, or the like. Usually during the preparation of a construct where various fragments are joined, the fragments, intermediate constructs and constructs will be carried on a cloning vector comprising a replication system functional in a prokaryotic host, e.g., E. coli and a marker for selection, e.g., biocide resistance, complementation to an auxotrophic host, etc. Other functional sequences may also be present, such as polylinkers, for ease of introduction and excision of the construct or portions thereof, or the like. A large number of cloning vectors are available such as pBR322, the pUC series, etc. These constructs may then be used for integration into the primary mammalian host.

In the case of the primary mammalian host, a replicating vector may be used. Usually, such vector will have a viral replication system, such as SV40, bovine papilloma virus, adenovirus, or the like. The linear DNA sequence vector may also have a selectable marker for identifying transfected cells. Selectable markers include the neo gene, allowing for selection with G418, the herpes tk gene for selection with HAT medium, the gpt gene with mycophenolic acid, complementation of an auxotrophic host, etc.

The vector may or may not be capable of stable maintenance in the host. Where the vector is capable of stable maintenance, the cells will be screened for homologous integration of the vector into the genome of the host, where various techniques for curing the cells may be employed. Where the vector is not capable of stable maintenance, for example, where a temperature sensitive replication system is employed, one may change the temperature from the permissive temperature to the non-permissive temperature, so that the cells may be cured of the vector. In this case, only those cells having integration of the construct comprising the amplifiable gene and, when present, the selectable marker, will be able to survive selection.

Where a selectable marker is present, one may select for the presence of the targeting construct by means of the selectable marker. Where the selectable marker is not present, one may select for the presence of the construct by the amplifiable gene. For the neo gene or the herpes tk gene, one could employ a medium for growth of the transformants of about 0.1-1 mg/ml of G418 or may use HAT medium, respectively. Where DHFR is the amplifiable gene, the selective medium may include from about 0.01-0.5 μM of methotrexate or be deficient in glycine-hypoxanthine-thymidine and have dialysed serum (GHT media).

The DNA can be introduced into the expression host by a variety of techniques that include calcium phosphate/DNA co-precipitates, microinjection of DNA into the nucleus, electroporation, yeast protoplast fusion with intact cells, transfection, polycations, e.g., polybrene, polyornithine, etc., or the like. The DNA may be single or double stranded DNA, linear or circular. The various techniques for transforming mammalian cells are well known (see Keown et al., Methods Enzymol. (1989); Keown et al., Methods Enzymol. 185:527-537 (1990); Mansour et al., Nature 336:348-352, (1988); all of which are herein incorporated by reference in their entirety).

(d) Insect Constructs and Transformed Insect Cells

The present invention also relates to an insect recombinant vectors comprising exogenous genetic material. The present invention also relates to an insect cell comprising an insect recombinant vector. The present invention also relates to methods for obtaining a recombinant insect host cell, comprising introducing into an insect cell exogenous genetic material.

The insect recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures and can bring about the expression of the nucleic acid sequence. The choice of a vector will typically depend on the compatibility of the vector with the insect host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the insect host. In addition, the insect vector may be an expression vector. Nucleic acid molecules can be suitably inserted into a replication vector for expression in the insect cell under a suitable promoter for insect cells. Many vectors are available for this purpose and selection of the appropriate vector will depend mainly on the size of the nucleic acid molecule to be inserted into the vector and the particular host cell to be transformed with the vector. Each vector contains various components depending on its function (amplification of DNA or expression of DNA) and the particular host cell with which it is compatible. The vector components for insect cell transformation generally include, but are not limited to, one or more of the following: a signal sequence, origin of replication, one or more marker genes and an inducible promoter.

The insect vector may be an autonomously replicating vector, i.e., a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any means for assuring self-replication. Alternatively, the vector may be one which, when introduced into the insect cell, is integrated into the genome and replicated together with the chromosome(s) into which it has been integrated. For integration, the vector may rely on the nucleic acid sequence of the vector for stable integration of the vector into the genome by homologous or nonhomologous recombination. Alternatively, the vector may contain additional nucleic acid sequences for directing integration by homologous recombination into the genome of the insect host. The additional nucleic acid sequences enable the vector to be integrated into the host cell genome at a precise location(s) in the chromosome(s). To increase the likelihood of integration at a precise location, there should be preferably two nucleic acid sequences which individually contain a sufficient number of nucleic acids, preferably 400 bp to 1500 bp, more preferably 800 bp to 1000 bp, which are highly homologous with the corresponding target sequence to enhance the probability of homologous recombination. These nucleic acid sequences may be any sequence that is homologous with a target sequence in the genome of the insect host cell and, furthermore, may be non-encoding or encoding sequences.

Baculovirus expression vectors (BEVs) have become important tools for the expression of foreign genes, both for basic research and for the production of proteins with direct clinical applications in human and veterinary medicine (Doerfler, Curr. Top. Microbiol. Immunol. 131:51-68 (1968); Luckow and Summers, Bio/Technology 6:47-55 (1988a); Miller, Annual Review of Microbiol. 42:177-199 (1988); Summers, Curr. Comm. Molecular Biology, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1988); all of which are herein incorporated by reference in their entirety). BEVs are recombinant insect viruses in which the coding sequence for a chosen foreign gene has been inserted behind a baculovirus promoter in place of the viral gene, e.g., polyhedrin (Smith and Summers, U.S. Pat. No. 4,745,051, the entirety of which is incorporated herein by reference).

The use of baculovirus vectors relies upon the host cells being derived from Lepidopteran insects such as Spodoptera frugiperda or Trichoplusia ni. The preferred Spodoptera frugiperda cell line is the cell line Sf9. The Spodoptera frugiperda Sf9 cell line was obtained from American Type Culture Collection (Manassas, Va.) and is assigned accession number ATCC CRL 1711 (Summers and Smith, A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Ag. Exper. Station Bulletin No. 1555 (1988), the entirety of which is herein incorporated by reference). Other insect cell systems, such as the silkworm B. mori may also be used.

The proteins expressed by the BEVs are, therefore, synthesized, modified and transported in host cells derived from Lepidopteran insects. Most of the genes that have been inserted and produced in the baculovirus expression vector system have been derived from vertebrate species. Other baculovirus genes in addition to the polyhedrin promoter may be employed to advantage in a baculovirus expression system. These include immediate-early (alpha), delayed-early (β), late (γ), or very late (delta), according to the phase of the viral infection during which they are expressed. The expression of these genes occurs sequentially, probably as the result of a “cascade” mechanism of transcriptional regulation. (Guarino and Summers, J. Virol. 57:563-571 (1986); Guarino and Summers, J. Virol. 61:2091-2099 (1987); Guarino and Summers, Virol. 162:444-451 (1988); all of which are herein incorporated by reference in their entirety).

Insect recombinant vectors are useful as intermediates for the infection or transformation of insect cell systems. For example, an insect recombinant vector containing a nucleic acid molecule encoding a baculovirus transcriptional promoter followed downstream by an insect signal DNA sequence is capable of directing the secretion of the desired biologically active protein from the insect cell. The vector may utilize a baculovirus transcriptional promoter region derived from any of the over 500 baculoviruses generally infecting insects, such as for example the Orders Lepidoptera, Diptera, Orthoptera, Coleoptera and Hymenoptera, including for example but not limited to the viral DNAs of Autographa californica MNPV, Bombyx mori NPV, Trichoplusia ni MNPV, Rachiplusia ou MNPV or Galleria mellonella MNPV, wherein said baculovirus transcriptional promoter is a baculovirus immediate-early gene IEI or IEN promoter; an immediate-early gene in combination with a baculovirus delayed-early gene promoter region selected from the group consisting of 39K and a HindIII-k fragment delayed-early gene; or a baculovirus late gene promoter. The immediate-early or delayed-early promoters can be enhanced with transcriptional enhancer elements. The insect signal DNA sequence may code for a signal peptide of a Lepidopteran adipokinetic hormone precursor or a signal peptide of the Manduca sexta adipokinetic hormone precursor (Summers, U.S. Pat. No. 5,155,037; the entirety of which is herein incorporated by reference). Other insect signal DNA sequences include a signal peptide of the Orthoptera Schistocerca gregaria locust adipokinetic hormone precurser and the Drosophila melanogaster cuticle genes CP1, CP2, CP3 or CP4 or for an insect signal peptide having substantially a similar chemical composition and function (Summers, U.S. Pat. No. 5,155,037).

Insect cells are distinctly different from animal cells. Insects have a unique life cycle and have distinct cellular properties such as the lack of intracellular plasminogen activators in which are present in vertebrate cells. Another difference is the high expression levels of protein products ranging from 1 to greater than 500 mg/liter and the ease at which cDNA can be cloned into cells (Frasier, In Vitro Cell. Dev. Biol. 25:225 (1989); Summers and Smith, In: A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Ag. Exper. Station Bulletin No. 1555 (1988), both of which are incorporated by reference in their entirety).

Recombinant protein expression in insect cells is achieved by viral infection or stable transformation. For viral infection, the desired gene is cloned into baculovirus at the site of the wild-type polyhedron gene (Webb and Summers, Technique 2:173 (1990); Bishop and Posse, Adv. Gene Technol. 1:55 (1990); both of which are incorporated by reference in their entirety). The polyhedron gene is a component of a protein coat in occlusions which encapsulate virus particles. Deletion or insertion in the polyhedron gene results the failure to form occlusion bodies. Occlusion negative viruses are morphologically different from occlusion positive viruses and enable one skilled in the art to identify and purify recombinant viruses.

The vectors of present invention preferably contain one or more selectable markers which permit easy selection of transformed cells. A selectable marker is a gene the product of which provides, for example biocide or viral resistance, resistance to heavy metals, prototrophy to auxotrophs and the like. Selection may be accomplished by co-transformation, e.g., as described in WO 91/17243, a nucleic acid sequence of the present invention may be operably linked to a suitable promoter sequence. The promoter sequence is a nucleic acid sequence which is recognized by the insect host cell for expression of the nucleic acid sequence. The promoter sequence contains transcription and translation control sequences which mediate the expression of the protein or fragment thereof. The promoter may be any nucleic acid sequence which shows transcriptional activity in the insect host cell of choice and may be obtained from genes encoding polypeptides either homologous or heterologous to the host cell.

For example, a nucleic acid molecule encoding a protein or fragment thereof may also be operably linked to a suitable leader sequence. A leader sequence is a nontranslated region of a mRNA which is important for translation by the fungal host. The leader sequence is operably linked to the 5′ terminus of the nucleic acid sequence encoding the protein or fragment thereof. The leader sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any leader sequence which is functional in the insect host cell of choice may be used in the present invention.

A polyadenylation sequence may also be operably linked to the 3′ terminus of the nucleic acid sequence of the present invention. The polyadenylation sequence is a sequence which when transcribed is recognized by the insect host to add polyadenosine residues to transcribed mRNA. The polyadenylation sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any polyadenylation sequence which is functional in the fungal host of choice may be used in the present invention.

To avoid the necessity of disrupting the cell to obtain the protein or fragment thereof and to minimize the amount of possible degradation of the expressed polypeptide within the cell, it is preferred that expression of the polypeptide gene gives rise to a product secreted outside the cell. To this end, the protein or fragment thereof of the present invention may be linked to a signal peptide linked to the amino terminus of the protein or fragment thereof. A signal peptide is an amino acid sequence which permits the secretion of the protein or fragment thereof from the insect host into the culture medium. The signal peptide may be native to the protein or fragment thereof of the invention or may be obtained from foreign sources. The 5′ end of the coding sequence of the nucleic acid sequence of the present invention may inherently contain a signal peptide coding region naturally linked in translation reading frame with the segment of the coding region which encodes the secreted protein or fragment thereof.

At present, a mode of achieving secretion of a foreign gene product in insect cells is by way of the foreign gene's native signal peptide. Because the foreign genes are usually from non-insect organisms, their signal sequences may be poorly recognized by insect cells and hence, levels of expression may be suboptimal. However, the efficiency of expression of foreign gene products seems to depend primarily on the characteristics of the foreign protein. On average, nuclear localized or non-structural proteins are most highly expressed, secreted proteins are intermediate and integral membrane proteins are the least expressed. One factor generally affecting the efficiency of the production of foreign gene products in a heterologous host system is the presence of native signal sequences (also termed presequences, targeting signals, or leader sequences) associated with the foreign gene. The signal sequence is generally coded by a DNA sequence immediately following (5′ to 3′) the translation start site of the desired foreign gene.

The expression dependence on the type of signal sequence associated with a gene product can be represented by the following example: If a foreign gene is inserted at a site downstream from the translational start site of the baculovirus polyhedrin gene so as to produce a fusion protein (containing the N-terminus of the polyhedrin structural gene), the fused gene is highly expressed. But less expression is achieved when a foreign gene is inserted in a baculovirus expression vector immediately following the transcriptional start site and totally replacing the polyhedrin structural gene.

Insertions into the region −50 to −1 significantly alter (reduce) steady state transcription which, in turn, reduces translation of the foreign gene product. Use of the pVL941 vector optimizes transcription of foreign genes to the level of the polyhedrin gene transcription. Even though the transcription of a foreign gene may be optimal, optimal translation may vary because of several factors involving processing: signal peptide recognition, mRNA and ribosome binding, glycosylation, disulfide bond formation, sugar processing, oligomerization, for example.

The properties of the insect signal peptide are expected to be more optimal for the efficiency of the translation process in insect cells than those from vertebrate proteins. This phenomenon can generally be explained by the fact that proteins secreted from cells are synthesized as precursor molecules containing hydrophobic N-terminal signal peptides. The signal peptides direct transport of the select protein to its target membrane and are then cleaved by a peptidase on the membrane, such as the endoplasmic reticulum, when the protein passes through it.

Another exemplary insect signal sequence is the sequence encoding for Drosophila cuticle proteins such as CP1, CP2, CP3 or CP4 (Summers, U.S. Pat. No. 5,278,050; the entirety of which is herein incorporated by reference). Most of a 9 kb region of the Drosophila genome containing genes for the cuticle proteins has been sequenced. Four of the five cuticle genes contains a signal peptide coding sequence interrupted by a short intervening sequence (about 60 base pairs) at a conserved site. Conserved sequences occur in the 5′ mRNA untranslated region, in the adjacent 35 base pairs of upstream flanking sequence and at −200 base pairs from the mRNA start position in each of the cuticle genes.

Standard methods of insect cell culture, cotransfection and preparation of plasmids are set forth in Summers and Smith (Summers and Smith, A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Agricultural Experiment Station Bulletin No. 1555, Texas A&M University (1987)). Procedures for the cultivation of viruses and cells are described in Volkman and Summers, J. Virol 19:820-832 (1975) and Volkman et al., J. Virol 19:820-832 (1976); both of which are herein incorporated by reference in their entirety.

(e) Bacterial Constructs and Transformed Bacterial Cells

The present invention also relates to a bacterial recombinant vector comprising exogenous genetic material. The present invention also relates to a bacteria cell comprising a bacterial recombinant vector. The present invention also relates to methods for obtaining a recombinant bacteria host cell, comprising introducing into a bacterial host cell exogenous genetic material.

The bacterial recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures. The choice of a vector will typically depend on the compatibility of the vector with the bacterial host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the bacterial host. In addition, the bacterial vector may be an expression vector. Nucleic acid molecules encoding protein homologues or fragments thereof can, for example, be suitably inserted into a replicable vector for expression in the bacterium under the control of a suitable promoter for bacteria. Many vectors are available for this purpose and selection of the appropriate vector will depend mainly on the size of the nucleic acid to be inserted into the vector and the particular host cell to be transformed with the vector. Each vector contains various components depending on its function (amplification of DNA or expression of DNA) and the particular host cell with which it is compatible. The vector components for bacterial transformation generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes and an inducible promoter.

In general, plasmid vectors containing replicon and control sequences that are derived from species compatible with the host cell are used in connection with bacterial hosts. The vector ordinarily carries a replication site, as well as marking sequences that are capable of providing phenotypic selection in transformed cells. For example, E. coli is typically transformed using pBR322, a plasmid derived from an E. coli species (see, e.g., Bolivar et al., Gene 2:95 (1977); the entirety of which is herein incorporated by reference). pBR322 contains genes for ampicillin and tetracycline resistance and thus provides easy means for identifying transformed cells. The pBR322 plasmid, or other microbial plasmid or phage, also generally contains, or is modified to contain, promoters that can be used by the microbial organism for expression of the selectable marker genes.

Nucleic acid molecules encoding protein or fragments thereof may be expressed not only directly, but also as a fusion with another polypeptide, preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature polypeptide. In general, the signal sequence may be a component of the vector, or it may be a part of the polypeptide DNA that is inserted into the vector. The heterologous signal sequence selected should be one that is recognized and processed (i.e., cleaved by a signal peptidase) by the host cell. For bacterial host cells that do not recognize and process the native polypeptide signal sequence, the signal sequence is substituted by a bacterial signal sequence selected, for example, from the group consisting of the alkaline phosphatase, penicillinase, lpp, or heat-stable enterotoxin II leaders.

Both expression and cloning vectors contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells. Generally, in cloning vectors this sequence is one that enables the vector to replicate independently of the host chromosomal DNA and includes origins of replication or autonomously replicating sequences. Such sequences are well known for a variety of bacteria. The origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria.

Expression and cloning vectors also generally contain a selection gene, also termed a selectable marker. This gene encodes a protein necessary for the survival or growth of transformed host cells grown in a selective culture medium. Host cells not transformed with the vector containing the selection gene will not survive in the culture medium. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli. One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous protein homologue or fragment thereof produce a protein conferring drug resistance and thus survive the selection regimen.

The expression vector for producing a protein or fragment thereof can also contains an inducible promoter that is recognized by the host bacterial organism and is operably linked to the nucleic acid encoding, for example, the nucleic acid molecule encoding the protein homologue or fragment thereof of interest. Inducible promoters suitable for use with bacterial hosts include the β-lactamase and lactose promoter systems (Chang et al., Nature 275:615 (1978); Goeddel et al., Nature 281:544 (1979); both of which are herein incorporated by reference in their entirety), the arabinose promoter system (Guzman et al., J. Bacteriol. 174:7716-7728 (1992); the entirety of which is herein incorporated by reference), alkaline phosphatase, a tryptophan (trp) promoter system (Goeddel, Nucleic Acids Res. 8:4057 (1980); EP 36,776; both of which are herein incorporated by reference in their entirety) and hybrid promoters such as the tac promoter (deBoer et al., Proc. Natl. Acad. Sci. (USA) 80:21-25 (1983); the entirety of which is herein incorporated by reference). However, other known bacterial inducible promoters are suitable (Siebenlist et al., Cell 20:269 (1980); the entirety of which is herein incorporated by reference).

Promoters for use in bacterial systems also generally contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding the polypeptide of interest. The promoter can be removed from the bacterial source DNA by restriction enzyme digestion and inserted into the vector containing the desired DNA.

Construction of suitable vectors containing one or more of the above-listed components employs standard ligation techniques. Isolated plasmids or DNA fragments are cleaved, tailored and re-ligated in the form desired to generate the plasmids required. Examples of available bacterial expression vectors include, but are not limited to, the multifunctional E. coli cloning and expression vectors such as Bluescript™ (Stratagene, La Jolla, Calif.), in which, for example, encoding an A. nidulans protein homologue or fragment thereof homologue, may be ligated into the vector in frame with sequences for the amino-terminal Met and the subsequent 7 residues of β-galactosidase so that a hybrid protein is produced; pIN vectors (Van Heeke and Schuster, J. Biol. Chem. 264:5503-5509 (1989), the entirety of which is herein incorporated by reference); and the like. pGEX vectors (Promega, Madison Wis. U.S.A.) may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins made in such systems are designed to include heparin, thrombin or factor XA protease cleavage sites so that the cloned polypeptide of interest can be released from the GST moiety at will.

Suitable host bacteria for a bacterial vector include archaebacteria and eubacteria, especially eubacteria and most preferably Enterobacteriaceae. Examples of useful bacteria include Escherichia, Enterobacter, Azotobacter, Erwinia, Bacillus, Pseudomonas, Klebsiella, Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla and Paracoccus. Suitable E. coli hosts include E. coli W3110 (American Type Culture Collection (ATCC) 27,325, Manassas, Va. U.S.A.), E. coli 294 (ATCC 31,446), E. coli B and E. coli X1776 (ATCC 31,537). These examples are illustrative rather than limiting. Mutant cells of any of the above-mentioned bacteria may also be employed. It is, of course, necessary to select the appropriate bacteria taking into consideration replicability of the replicon in the cells of a bacterium. For example, E. coli, Serratia, or Salmonella species can be suitably used as the host when well known plasmids such as pBR322, pBR325, pACYC177, or pKN410 are used to supply the replicon. E. coli strain W3110 is a preferred host or parent host because it is a common host strain for recombinant DNA product fermentations. Preferably, the host cell should secrete minimal amounts of proteolytic enzymes.

Host cells are transfected and preferably transformed with the above-described vectors and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.

Numerous methods of transfection are known to the ordinarily skilled artisan, for example, calcium phosphate and electroporation. Depending on the host cell used, transformation is done using standard techniques appropriate to such cells. The calcium treatment employing calcium chloride, as described in section 1.82 of Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, (1989), is generally used for bacterial cells that contain substantial cell-wall barriers. Another method for transformation employs polyethylene glycol/DMSO, as described in Chung and Miller (Chung and Miller, Nucleic Acids Res. 16:3580 (1988); the entirety of which is herein incorporated by reference). Yet another method is the use of the technique termed electroporation.

Bacterial cells used to produce the polypeptide of interest for purposes of this invention are cultured in suitable media in which the promoters for the nucleic acid encoding the heterologous polypeptide can be artificially induced as described generally, e.g., in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, (1989). Examples of suitable media are given in U.S. Pat. Nos. 5,304,472 and 5,342,763; both of which are incorporated by reference in their entirety.

In addition to the above discussed procedures, practitioners are familiar with the standard resource materials which describe specific conditions and procedures for the construction, manipulation and isolation of macromolecules (e.g., DNA molecules, plasmids, etc.), generation of recombinant organisms and the screening and isolating of clones, (see for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989); Mailga et al., Methods in Plant Molecular Biology, Cold Spring Harbor Press (1995), the entirety of which is herein incorporated by reference; Birren et al., Genome Analysis: Analyzing DNA, 1, Cold Spring Harbor, N.Y., the entirety of which is herein incorporated by reference).

(f) Computer Readable Media

The nucleotide sequence provided in SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof, or complement thereof, or a nucleotide sequence at least 90% identical, preferably 95%, identical even more preferably 99% or 100% identical to the sequence provided in SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof, or complement thereof, can be “provided” in a variety of mediums to facilitate use. Such a medium can also provide a subset thereof in a form that allows a skilled artisan to examine the sequences.

A preferred subset of nucleotide sequences are those nucleic acid sequences that encode a maize or soybean methionine adenosyltransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean S-adenosylmethionine decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean aspartate kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean aspartate-semialdehyde dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean cystathionine β-lyase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cytsathionine γ-lyase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complement thereof or fragment of either.

A further preferred subset of nucleic acid sequences is where the subset of sequences is two proteins or fragments thereof, more preferably three proteins or fragments thereof and even more preferable four proteins or fragments thereof, these nucleic acid sequences are selected from the group that comprises a maize or soybean methionine adenosyltransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean S-adenosylmethionine decarboxylase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean aspartate kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean aspartate-semialdehyde dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean cystathionine β-lyase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or soybean 5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean adenosylhomocysteinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cystathionine β-synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean cytsathionine γ-lyase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complement thereof or fragment of either.

In one application of this embodiment, a nucleotide sequence of the present invention can be recorded on computer readable media. As used herein, “computer readable media” refers to any medium that can be read and accessed directly by a computer. Such media include, but are not limited to: magnetic storage media, such as floppy discs, hard disc, storage medium and magnetic tape: optical storage media such as CD-ROM; electrical storage media such as RAM and ROM; and hybrids of these categories such as magnetic/optical storage media. A skilled artisan can readily appreciate how any of the presently known computer readable mediums can be used to create a manufacture comprising computer readable medium having recorded thereon a nucleotide sequence of the present invention.

As used herein, “recorded” refers to a process for storing information on computer readable medium. A skilled artisan can readily adopt any of the presently known methods for recording information on computer readable medium to generate media comprising the nucleotide sequence information of the present invention. A variety of data storage structures are available to a skilled artisan for creating a computer readable medium having recorded thereon a nucleotide sequence of the present invention. The choice of the data storage structure will generally be based on the means chosen to access the stored information. In addition, a variety of data processor programs and formats can be used to store the nucleotide sequence information of the present invention on computer readable medium. The sequence information can be represented in a word processing text file, formatted in commercially-available software such as WordPerfect and Microsoft Word, or represented in the form of an ASCII file, stored in a database application, such as DB2, Sybase, Oracle, or the like. A skilled artisan can readily adapt any number of data processor structuring formats (e.g. text file or database) in order to obtain computer readable medium having recorded thereon the nucleotide sequence information of the present invention.

By providing one or more of nucleotide sequences of the present invention, a skilled artisan can routinely access the sequence information for a variety of purposes. Computer software is publicly available which allows a skilled artisan to access sequence information provided in a computer readable medium. The examples which follow demonstrate how software which implements the BLAST (Altschul et al., J. Mol. Biol. 215:403-410 (1990), the entirety of which is herein incorporated by reference) and BLAZE (Brutlag et al., Comp. Chem. 17:203-207 (1993), the entirety of which is herein incorporated by reference) search algorithms on a Sybase system can be used to identify open reading frames (ORFs) within the genome that contain homology to ORFs or proteins from other organisms. Such ORFs are protein-encoding fragments within the sequences of the present invention and are useful in producing commercially important proteins such as enzymes used in amino acid biosynthesis, metabolism, transcription, translation, RNA processing, nucleic acid and a protein degradation, protein modification and DNA replication, restriction, modification, recombination and repair.

The present invention further provides systems, particularly computer-based systems, which contain the sequence information described herein. Such systems are designed to identify commercially important fragments of the nucleic acid molecule of the present invention. As used herein, “a computer-based system” refers to the hardware means, software means and data storage means used to analyze the nucleotide sequence information of the present invention. The minimum hardware means of the computer-based systems of the present invention comprises a central processing unit (CPU), input means, output means and data storage means. A skilled artisan can readily appreciate that any one of the currently available computer-based system are suitable for use in the present invention.

As indicated above, the computer-based systems of the present invention comprise a data storage means having stored therein a nucleotide sequence of the present invention and the necessary hardware means and software means for supporting and implementing a search means. As used herein, “data storage means” refers to memory that can store nucleotide sequence information of the present invention, or a memory access means which can access manufactures having recorded thereon the nucleotide sequence information of the present invention. As used herein, “search means” refers to one or more programs which are implemented on the computer-based system to compare a target sequence or target structural motif with the sequence information stored within the data storage means. Search means are used to identify fragments or regions of the sequence of the present invention that match a particular target sequence or target motif. A variety of known algorithms are disclosed publicly and a variety of commercially available software for conducting search means are available can be used in the computer-based systems of the present invention. Examples of such software include, but are not limited to, MacPattern (EMBL), BLASTIN and BLASTIX (NCBIA). One of the available algorithms or implementing software packages for conducting homology searches can be adapted for use in the present computer-based systems.

The most preferred sequence length of a target sequence is from about 10 to 100 amino acids or from about 30 to 300 nucleotide residues. However, it is well recognized that during searches for commercially important fragments of the nucleic acid molecules of the present invention, such as sequence fragments involved in gene expression and protein processing, may be of shorter length.

As used herein, “a target structural motif,” or “target motif,” refers to any rationally selected sequence or combination of sequences in which the sequences the sequence(s) are chosen based on a three-dimensional configuration which is formed upon the folding of the target motif. There are a variety of target motifs known in the art. Protein target motifs include, but are not limited to, enzymatic active sites and signal sequences. Nucleic acid target motifs include, but are not limited to, promoter sequences, cis elements, hairpin structures and inducible expression elements (protein binding sequences).

Thus, the present invention further provides an input means for receiving a target sequence, a data storage means for storing the target sequences of the present invention sequence identified using a search means as described above and an output means for outputting the identified homologous sequences. A variety of structural formats for the input and output means can be used to input and output information in the computer-based systems of the present invention. A preferred format for an output means ranks fragments of the sequence of the present invention by varying degrees of homology to the target sequence or target motif. Such presentation provides a skilled artisan with a ranking of sequences which contain various amounts of the target sequence or target motif and identifies the degree of homology contained in the identified fragment.

A variety of comparing means can be used to compare a target sequence or target motif with the data storage means to identify sequence fragments sequence of the present invention. For example, implementing software which implement the BLAST and BLAZE algorithms (Altschul et al., J. Mol. Biol. 215:403-410 (1990)) can be used to identify open frames within the nucleic acid molecules of the present invention. A skilled artisan can readily recognize that any one of the publicly available homology search programs can be used as the search means for the computer-based systems of the present invention.

Having now generally described the invention, the same will be more readily understood through reference to the following examples which are provided by way of illustration and are not intended to be limiting of the present invention, unless specified.

Example 1

The MONN01 cDNA library is a normalized library generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON001 cDNA library is generated from maize (B73, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) immature tassels at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V6 stage. At that stage the tassel is an immature tassel of about 2-3 cm in length. The tassels are removed and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON003 library is generated from maize (B73 x Mo17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) roots at the V6 developmental stage. Seeds are planted at a depth of approximately 3 cm in coil into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth, the seedlings are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and approximately 3 times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting at a concentration of 150 ppm N. Two to three times during the life time of the plant from transplanting to flowering a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in approximately 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6 leaf development stage. The root system is cut from maize plant and washed with water to free it from the soil. The tissue is then immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON004 cDNA library is generated from maize (B73 x Mo17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON005 cDNA library is generated from maize (B73 x Mo17, Illinois Foundation Seeds, Champaign Ill., U.S.A.) root tissue at the V6 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen and the harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON006 cDNA library is generated from maize (B73 x Mo17, Illinois Foundation Seeds, Champaign Ill., U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON007 cDNA library is generated from the primary root tissue of 5 day old maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). After germination, the trays, along with the moist paper, are moved to a greenhouse where the maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles for approximately 5 days. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. The primary root tissue is collected when the seedlings are 5 days old. At this stage, the primary root (radicle) is pushed through the coleorhiza which itself is pushed through the seed coat. The primary root, which is about 2-3 cm long, is cut and immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON008 cDNA library is generated from the primary shoot (coleoptile 2-3 cm) of maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings which are approximately 5 days old. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). Then the trays containing the seeds are moved to a greenhouse at 15 hr daytime/9 hr nighttime cycles and grown until they are 5 days post germination. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Tissue is collected when the seedlings are 5 days old. At this stage, the primary shoot (coleoptile) is pushed through the seed coat and is about 2-3 cm long. The coleoptile is dissected away from the rest of the seedling, immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON009 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaves at the 8 leaf stage (V8 plant development stage). Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 8-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical, are cut at the base of the leaves. The leaves are then pooled and then immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON010 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) root tissue at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the V8 development stage. The root system is cut from this mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON011 cDNA library is generated from undeveloped maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaf at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The second youngest leaf which is at the base of the apical leaf of V6 stage maize plant is cut at the base and immediately transferred to liquid nitrogen containers in which the leaf is crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON012 cDNA library is generated from 2 day post germination maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). Then the trays containing the seeds are moved to the greenhouse and grown at 15 hr daytime/9 hr nighttime cycles until 2 days post germination. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Tissue is collected when the seedlings are 2 days old. At the two day stage, the coleorhiza is pushed through the seed coat and the primary root (the radicle) is pierced the coleorhiza but is barely visible. Also, at this two day stage, the coleoptile is just emerging from the seed coat. The 2 days post germination seedlings are then immersed in liquid nitrogen and crushed. The harvested tissue is stored at −80° C. until preparation of total RNA.

The SATMON013 cDNA library is generated from apical maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) meristem founder at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, the plant is at the 4 leaf stage. The lead at the apex of the V4 stage maize plant is referred to as the meristem founder. This apical meristem founder is cut, immediately frozen in liquid nitrogen and crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON014 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) endosperm fourteen days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the maize plant ear shoots are ready for fertilization. At this stage, the ear shoots are enclosed in a paper bag before silk emergence to withhold the pollen. The ear shoots are pollinated and 14 days after pollination, the ears are pulled out and then the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the endosperms are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON016 library is a maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) sheath library collected at the V8 developmental stage. Seeds are planted in a depth of approximately 3 cm in solid into 2-3 inch pots containing Metro growing medium. After 2-3 weeks growth, they are transplanted into 10″ pots containing the same. Plants are watered daily before transplantation and approximately the times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting, at a strength of 150 ppm N. Two to three times during the life time of the plant from transplanting to flowering, a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 w sodium vapor lamps. When the maize plants are at the V8 stage the 5^(th) and 6^(th) leaves from the bottom exhibit fully developed leaf blades. At the base of these leaves, the ligule is differentiated and the leaf blade is joined to the sheath. The sheath is dissected away from the base of the leaf then the sheath is frozen in liquid nitrogen and crushed. The tissue is then stored at −80° C. until RNA preparation.

The SATMON017 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) embryo seventeen days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth the seeds are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergence to withhold the pollen. The ear shoots are fertilized and 21 days after pollination, the ears are pulled out and the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the embryos are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON019 (Lib3054) cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) culm (stem) at the V8 developmental stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. When the maize plant is at the V8 stage, the 5th and 6th leaves from the bottom have fully developed leaf blades. The region between the nodes of the 5th and the sixth leaves from the bottom is the region of the stem that is collected. The leaves are pulled out and the sheath is also torn away from the stem. This stem tissue is completely free of any leaf and sheath tissue. The stem tissue is then frozen in liquid nitrogen and stored at −80° C. until RNA preparation.

The SATMON020 cDNA library is from a maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) Hill Type II-Initiated Callus. Petri plates containing approximately 25 ml of Type II initiation media are prepared. This medium contains N6 salts and vitamins, 3% sucrose, 2.3 g/liter proline 0.1 g/liter enzymatic casein hydrolysate, 2 mg/liter 2,4-dichloro phenoxy-acetic acid (2,4, D), 15.3 mg/liter AgNO₃ and 0.8% bacto agar and is adjusted to pH 6.0 before autoclaving. At 9-11 days after pollination, an ear with immature embryos measuring approximately 1-2 mm in length is chosen. The husks and silks are removed and then the ear is broken into halves and placed in an autoclaved solution of Clorox/TWEEN 20 sterilizing solution. Then the ear is rinsed with deionized water. Then each embryo is extracted from the kernel. Intact embryos are placed in contact with the medium, scutellar side up). Multiple embryos are plated on each plate and the plates are incubated in the dark at 25° C. Type II calluses are friable, can be subcultured with a spatula, frequently regenerate via somatic embryogenesis and are relatively undifferentiated. As seen in the microscope, the Tape II calluses show color ranging from translucent to light yellow and heterogeneity on with respect to embryoid structure as well as stage of embryoid development. Once Type II callus are formed, the calluses is transferred to type II callus maintenance medium without AgNO₃. Every 7-10 days, the callus is subcultured. About 4 weeks after embryo isolation the callus is removed from the plates and then frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The SATMON021 cDNA library is generated from the immature maize (DK604, Dekalb Genetics, Dekalb Ill., U.S.A.) tassel at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. As the maize plant enters the V8 stage, tassels which are 15-20 cm in length are collected and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The SATMON022 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) ear (growing silks) at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Zea mays plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the plant is in the V8 stage. At this stage, some immature ear shoots are visible. The immature ear shoots (approximately 1 cm in length) are pulled out, frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON23 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) ear (growing silk) at the V8 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. When the tissue is harvested at the V8 stage, the length of the ear that is harvested is about 10-15 cm and the silks are just exposed (approximately 1 inch). The ear along with the silks is frozen in liquid nitrogen and then the tissue is stored at −80° C. until RNA preparation.

The SATMON024 cDNA library is generated from the immature maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) tassel at the V9 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. As a maize plant enters the V9 stage, the tassel is rapidly developing and a 37 cm tassel along with the glume, anthers and pollen is collected and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The SATMON025 cDNA library is from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) Hill Type II-Regenerated Callus. Type II callus is grown in initiation media as described for SATMON020 and then the embryoids on the surface of the Type II callus are allowed to mature and germinate. The 1-2 gm fresh weight of the soft friable type callus containing numerous embryoids are transferred to 100×15 mm petri plates containing 25 ml of regeneration media. Regeneration media consists of Murashige and Skoog (MS) basal salts, modified White's vitamins (0.2 g/liter glycine and 0.5 g/liter myo-inositol and 0.8% bacto agar (6SMS0D)). The plates are then placed in the dark after covering with parafilm. After 1 week, the plates are moved to a lighted growth chamber with 16 hr light and 8 hr dark photoperiod. Three weeks after plating the Type II callus to 6SMS0D, the callus exhibit shoot formation. The callus and the shoots are transferred to fresh 6SMS0D plates for another 2 weeks. The callus and the shoots are then transferred to petri plates with reduced sucrose (3SMSOD). Upon distinct formation of a root and shoot, the newly developed green plants are then removed out with a spatula and frozen in liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON026 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) juvenile/adult shift leaves at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plants are at the 8-leaf development stage. Leaves are founded sequentially around the meristem over weeks of time and the older, more juvenile leaves arise earlier and in a more basal position than the younger, more adult leaves, which are in a more apical position. In a V8 plant, some leaves which are in the middle portion of the plant exhibit characteristics of both juvenile as well as adult leaves. They exhibit a yellowing color but also exhibit, in part, a green color. These leaves are termed juvenile/adult shift leaves. The juvenile/adult shift leaves (the 4th, 5th leaves from the bottom) are cut at the base, pooled and transferred to liquid nitrogen in which they are then crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON027 cDNA library is generated from 6 day maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaves. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Zea mays plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, when the plant is at the 8-leaf stage, water is held back for six days. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical, are all cut at the base of the leaves. All the leaves exhibit significant wilting. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are then crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON028 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) roots at the V8 developmental stage that are subject to six days water stress. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, when the plant is at the 8-leaf stage, water is held back for six days. The root system is cut, shaken and washed to remove soil. Root tissue is then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are then crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON029 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings at the etiolated stage. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark for 4 days at approximately 70° F. Tissue is collected when the seedlings are 4 days old. By 4 days, the primary root has penetrated the coleorhiza and is about 4-5 cm and the secondary lateral roots have also made their appearance. The coleoptile has also pushed through the seed coat and is about 4-5 cm long. The seedlings are frozen in liquid nitrogen and crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON030 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) root tissue at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth, they are transplanted into 10 inch pots containing the same. Plants are watered daily before transplantation and approximately 3 times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting, at a strength of 150 ppm N. Two to three times during the life time of the plant, from transplanting to flowering, a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 sodium vapor lamps. Tissue is collected when the maize plant is at the 4 leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is then immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON031 cDNA library is generated from the maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaf tissue at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 4-leaf development stage. The third leaf from the bottom is cut at the base and immediately frozen in liquid nitrogen and crushed. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON033 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) embryo tissue 13 days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of the maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergent to withhold the pollen. The ear shoots are pollinated and 13 days after pollination, the ears are pulled out and then the kernels are plucked cut of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the embryos are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON034 cDNA library is generated from cold stressed maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept on at 10° C. for 7 days. After 7 days, the temperature is shifted to 15° C. for one day until germination of the seed. Tissue is collected once the seedlings are 1 day old. At this point, the coleorhiza has just pushed out of the seed coat and the primary root is just making its appearance. The coleoptile has not yet pushed completely through the seed coat and is also just making its appearance. These 1 day old cold stressed seedlings are frozen in liquid nitrogen and crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON˜001 (Lib36, Lib83, Lib84) cDNA library is generated from maize leaves at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V8 stage. The older more juvenile leaves in a basal position was well as the younger more adult leaves which are more apical are all cut at the base, pooled and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMONN01 cDNA library is generated from maize (B73, Illinois Foundation Seeds, Champaign, Illinois, U.S.A.) normalized immature tassels at the V6 plant development stage normalized tissue. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the night time temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V6 stage. At that stage the tassel is an immature tassel of about 2-3 cm in length. The tassels are removed and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M280 streptavidin (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The SATMONN04 cDNA library is generated from maize (B73 x Mo 17, Illinois Foundation Seeds, Champaign, Illinois, U.S.A.) normalized total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately SOT and the night time temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M280 streptavidin (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The SATMONN05 cDNA library is generated from maize (B73 x Mo 17, Illinois Foundation Seeds, Champaign, Illinois, U.S.A.) normalized root tissue at the V6 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant. from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the night time temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen and the harvested tissue is then stored at −80° C. until RNA preparation. The single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M280 streptavidin (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The SATMONN06 cDNA library is generated from maize (B73 x Mo17, illinois Foundation Seeds, Champaign Illinois, U.S.A.) normalized total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the night time temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M280 streptavidin (DYNA13EADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The CMZ029 (SATMON036) cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) endosperm 22 days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of the maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergent to withhold the pollen. The ear shoots are pollinated and 22 days after pollination, the ears are pulled out and then the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the alurone layer is removed. After dissection, the endosperms are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The CMz030 (Lib143) cDNA library is generated from maize seedling tissue two days post germination. Seeds are planted on a moist filter paper on a covered try that is keep in the dark until germination. The trays are then moved to the bench top at 15 hr daytime/9 hr nighttime cycles for 2 days post-germination. The day time temperature is 80° F. and the nighttime temperature is 70° F. Tissue is collected when the seedlings are 2 days old. At this stage, the colehrhiza has pushed through the seed coat and the primary root (the radicle) is just piercing the colehrhiza and is barely visible. The seedlings are placed at 42° C. for 1 hour. Following the heat shock treatment, the seedlings are immersed in liquid nitrogen and crushed. The harvested tissue is stored at −80° until RNA preparation.

The CMz031 (Lib 148) cDNA library is generated from maize pollen tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag to withhold pollen. Twenty-one days after pollination, prior to removing the ears, the paper bag is shaken to collect the mature pollen. The mature pollen is immediately frozen in liquid nitrogen containers and the pollen is crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz033 (Lib189) cDNA library is generated from maize pooled leaf tissue. Samples are harvested from open pollinated plants. Tissue is collected from maize leaves at the anthesis stage. The leaves are collect from 10-12 plants and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz034 (Lib3060) cDNA library is generated from maize mature tissue at 40 days post pollination plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from leaves located two leaves below the ear leaf. This sample represents those genes expressed during onset and early stages of leaf senescence. The leaves are pooled and immediately transferred to liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz036 (Lib3062) cDNA library is generated from maize husk tissue at the 8 week old plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from 8 week old plants. The husk is separated from the ear and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz037 (Lib3059) cDNA library is generated from maize pooled kernal at 12-15 days after pollienation plant development stage. Sample were collected from field grown material. Whole kernals from hand pollinated (control pollination) are harvested as whole ears and immediately frozen on dry ice. Kernels from 10-12 ears were pooled and ground together in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz039 (Lib3066) cDNA library is generated from maize immature anther tissue at the 7 week old immature tassel stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 7 week old immature tassel stage. At this stage, prior to anthesis, the immature anthers are green and enclosed in the staminate spikelet. The developing anthers are dissected away from the 7 week old immature tassel and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz040 (Lib3067) cDNA library is generated from maize kernel tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag before silk emergence to withhold pollen. Five to eight days after controlled pollination. The ears are pulled and the kernels removed. The kernels are immediately frozen in liquid nitrogen. The harvested kernels tissue is then stored at −80° C. until RNA preparation. This sample represents gene expressed in early kernel development, during periods of cell division, amyloplast biogenesis and early carbon flow across the material to filial tissue.

The CMz041 (Lib3068) cDNA library is generated from maize pollen germinating silk tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants when the ear shoots are ready for fertilization at the silk emergence stage. The emerging silks are pollinated with an excess of pollen under controlled pollination conditions in the green house. Eighteen hours after pollination the silks are removed from the ears and immediately frozen in liquid nitrogen containers. This sample represents genes expressed in both pollen and silk tissue early in pollination. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz042 (Lib3069) cDNA library is generated from maize ear tissue excessively pollinated at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants and the ear shoots which are ready for fertilization are at the silk emergence stage. The immature ears are pollinated with an excess of pollen under controlled pollination conditions. Eighteen hours post-pollination, the ears are removed and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz044 (Lib3075) cDNA library is generated from maize microspore tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from immature anthers from 7 week old tassels. The immature anthers are first dissected from the 7 week old tassel with a scalpel on a glass slide covered with water. The microspores (immature pollen) are released into the water and are recovered by centrifugation. The microspore suspension is immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz045 (Lib3076) cDNA library is generated from maize immature ear megaspore tissue. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from immature ear (megaspore) obtained from 7 week old plants. The immature ears are harvested from the 7 week old plants and are approximately 2.5 to 3 cm in length. The kernels are removed from the cob immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz047 (Lib3078) cDNA library is generated from maize CO₂ treated high-exposure shoot tissue at the V10+ plant development stage. RX601 maize seeds are sterilized for i minute with a 10% clorox solution. The seeds are rolled in germination paper, and germinated in 0.5 mM calcium sulfate solution for two days ate 30° C. The seedlings are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium at a rate of 2-3 seedlings per pot. Twenty pots are placed into a high CO₂ environment (approximately 1000 ppm CO₂). Twenty plants were grown under ambient greenhouse CO₂ (approximately 450 ppm CO₂). Plants are watered daily before transplantation and three times a week after transplantation. Peters 20-20-20 fertilizer is also lightly applied. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. At ten days post planting, the shoots from both atmosphere are frozen in liquid nitrogen and lightly ground. The roots are washed in deionized water to remove the support media and the tissue is immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz048 (Lib3079) cDNA library is generated from maize basal endosperm transfer layer tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ maize plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag prior to silk emergence, to withhold the pollen. Kernels are harvested at 12 days post-pollination and placed on wet ice for dissection. The kernels are cross sectioned laterally, dissecting just above the pedicel region, including 1-2 mm of the lower endosperm and the basal endosperm transfer region. The pedicel and lower endosperm region containing the basal endosperm transfer layer is pooled and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz049 (Lib3088) cDNA library is generated from maize immature anther tissue at the 7 week old immature tassel stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 7 week old immature tassel stage. At this stage, prior to anthesis, the immature anthers are green and enclosed in the staminate spikelet. The developing anthers are dissected away from the 7 week old immature tassel and immediately transferred to liquid nitrogen container. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz050 (Lib3114) cDNA library is generated from maize silk tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is beyond the 10-leaf development stage and the ear shoots are approximately 15-20 cm in length. The ears are pulled and silks are separated from the ears and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON001 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) total leaf tissue at the V4 plant development stage. Leaf tissue from 38, field grown V4 stage plants is harvested from the 4^(th) node. Leaf tissue is removed from the plants and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON002 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue at the V4 plant development stage. Root tissue from 76, field grown V4 stage plants is harvested. The root systems is cut from the soybean plant and washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON003 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotyl axis tissue harvested 2 day post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 2 days after the start of imbibition. The 2 days after imbibition samples are separated into 3 collections after removal of any adhering seed coat. At the 2 day stage, the hypocotyl axis is emerging from the soil. A few seedlings have cracked the soil surface and exhibited slight greening of the exposed cotyledons. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON004 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledon tissue harvested 2 day post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 2 days after the start of imbibition. The 2 days after imbibition samples are separated into 3 collections after removal of any adhering seed coat. At the 2 day stage, the hypocotyl axis is emerging from the soil. A few seedlings have cracked the soil surface and exhibited slight greening of the exposed cotyledons. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON005 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotyl axis tissue harvested 6 hour post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 6 hours after the start of imbibition. The 6 hours after imbibition samples are separated into 3 collections after removal of any adhering seed coat. The 6 hours after imbibition sample is collected over the course of approximately 2 hours starting at 6 hours post imbibition. At the 6 hours after imbibition stage, not all cotyledons have become fully hydrated and germination, or radicle protrusion, has not occurred. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON006 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledons tissue harvest 6 hour post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 6 hours after imbibition. The 6 hours after imbibition samples are separated into 3 collections after removal of any adhering seed coat. The 6 hours after imbibition sample is collected over the course of approximately 2 hours starting at 6 hours post-imbibition. At the 6 hours after imbibition, not all cotyledons have become fully hydrated and germination or radicle protrusion, have not occurred. The seedlings are washed in water to remove soil, cotyledon harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON007 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 and 35 days post-flowering. Seed pods from field grown plants are harvested 25 and 35 days after flowering and the seeds extracted from the pods. Approximately 4.4 g and 19.3 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON008 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested from 25 and 35 days post-flowering plants. Total leaf tissue is harvested from field grown plants. Approximately 19 g and 29 g of leaves are harvested from the fourth node of the plant 25 and 35 days post-flowering and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON009 cDNA library is generated from soybean cutlivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) pod and seed tissue harvested 15 days post-flowering. Pods from field grown plants are harvested 15 days post-flowering. Approximately 3 g of pod tissue is harvested and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON010 cDNA library is generated from soybean cultivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) seed tissue harvested 40 days post-flowering. Pods from field grown plants are harvested 40 days post-flowering. Pods and seeds are separated, approximately 19 g of seed tissue is harvested and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON011 cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4′ node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON012 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue. Leaves from field grown plants are harvested from the fourth node 15 days post-flowering. Approximately 12 g of leaves are harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON013 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root and nodule tissue. Approximately, 28 g of root tissue from field grown plants is harvested 15 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON014 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 and 35 days after flowering. Seed pods from field grown plants are harvested 15 days after flowering and the seeds extracted from the pods. Approximately 5 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON015 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 45 and 55 days post-flowering. Seed pods from field grown plants are harvested 45 and 55 days after flowering and the seeds extracted from the pods. Approximately 19 g and 31 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON016 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue. Approximately, 61 g and 38 g of root tissue from field grown plants is harvested 25 and 35 days post-flowering is harvested. The root system is cut from the soybean plant and washed with water to free it from the soil. The tissue is placed in 14 ml polystyrene tubes and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON017 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue. Approximately 28 g of root tissue from field grown plants is harvested 45 and 55 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON018 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested 45 and 55 days post-flowering. Leaves from field grown plants are harvested 45 and 55 days after flowering from the fourth node. Approximately 27 g and 33 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON019 cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) root tissue. Roots are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 50 g and 56 g of roots are harvested from each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON020 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 65 and 75 days post-flowering. Seed pods from field grown plants are harvested 45 and 55 days after flowering and the seeds extracted from the pods. Approximately 14 g and 31 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON021 cDNA library is generated from Soybean Cyst Nematode-resistant soybean cultivar Hartwig (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) root tissue. Plants are grown in tissue culture at room temperature. At approximately 6 weeks post-germination, the plants are exposed to sterilized Soybean Cyst Nematode eggs. Infection is then allowed to progress for 10 days. After the 10 day infection process, the tissue is harvested. Agar from the culture medium and nematodes are removed and the root tissue is immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON022 (Lib3030) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) partially opened flower tissue. Partially to fully opened flower tissue is harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. A total of 3 g of flower tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON023 cDNA library is generated from soybean genotype BW211S Null (Tohoku University, Morioka, Japan) seed tissue harvested 15 and 40 days post-flowering. Seed pods from field grown plants are harvested 15 and 40 days post-flowering and the seeds extracted from the pods. Approximately 0.7 g and 14.2 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON024 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) internode-2 tissue harvested 18 days post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. The plants are grown in a greenhouse for 18 days after the start of imbibition at ambient temperature. Soil is checked and watered daily to maintain even moisture conditions. Stem tissue is harvested 18 days after the start of imbibition. The samples are divided into hypocotyl and internodes 1 through 5. The fifth internode contains some leaf bud material. Approximately 3 g of each sample is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON025 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested 65 days post-flowering. Leaves are harvested from the fourth node of field grown plants 65 days post-flowering. Approximately 18.4 g of leaf tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

SOYMON026 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue harvested 65 and 75 days post-flowering. Approximately 27 g and 40 g of root tissue from field grown plants is harvested 65 and 75 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON027 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 days post-flowering. Seed pods from field grown plants are harvested 25 days post-flowering and the seeds extracted from the pods. Approximately 17 g of seeds are harvested from the seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON028 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought-stressed root tissue. The plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of development, water is withheld from half of the plant collection (drought stressed population). After 3 days, half of the plants from the drought stressed condition and half of the plants from the control population are harvested. After another 3 days (6 days post drought induction) the remaining plants are harvested. A total of 27 g and 40 g of root tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON029 cDNA library is generated from Soybean Cyst Nematode-resistant soybean cultivar PI07354 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) root tissue. Late fall to early winter greenhouse grown plants are exposed to Soybean Cyst Nematode eggs. At 10 days post-infection, the plants are uprooted, rinsed briefly and the roots frozen in liquid nitrogen. Approximately 20 grams of root tissue is harvested from the infected plants. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON030 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) flower bud tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Flower buds are removed from the plant at the pedicel. A total of 100 mg of flower buds are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON031 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) carpel and stamen tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Flower buds are removed from the plant at the pedicel. Flowers are dissected to separate petals, sepals and reproductive structures (carpels and stamens). A total of 300 mg of carpel and stamen tissue are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON032 cDNA library is prepared from the Asgrow cultivar A4922 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dry soybean seed meristem tissue. Surface sterilized seeds are germinated in liquid media for 24 hours. The seed axis is then excised from the barely germinating seed, placed on tissue culture media and incubated overnight at 20° C. in the dark. The supportive tissue is removed from the explant prior to harvest. Approximately 570 mg of tissue is harvested and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON033 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heat-shocked seedling tissue without cotyledons. Seeds are imbibed and germinated in vermiculite for 2 days under constant illumination. After 48 hours, the seedlings are transferred to an incubator set at 40° C. under constant illumination. After 30, 60 and 180 minutes seedlings are harvested and dissected. A portion of the seedling consisting of the root, hypocotyl and apical hook is frozen in liquid nitrogen and stored at −80° C. The seedlings after 2 days of imbibition are beginning to emerge from the vermiculite surface. The apical hooks are dark green in appearance. Total RNA and poly A⁺ RNA is prepared from equal amounts of pooled tissue.

The SOYMON034 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) cold-shocked seedling tissue without cotyledons. Seeds are imbibed and germinated in vermiculite for 2 days under constant illumination. After 48 hours, the seedlings are transferred to a cold room set at 5° C. under constant illumination. After 30, 60 and 180 minutes seedlings are harvested and dissected. A portion of the seedling consisting of the root, hypocotyl and apical hook is frozen in liquid nitrogen and stored at −80° C. The seedlings after 2 days of imbibition are beginning to emerge from the vermiculite surface. The apical hooks are dark green in appearance.

The SOYMON035 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed coat tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are harvested from mid to nearly full maturation (seed coats are not yellowing). The entire embryo proper is removed from the seed coat sample and the seed coat tissue are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON036 cDNA library is generated from soybean cultivars PI171451, PI227687 and PI229358 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) insect challenged leaves. Plants from each of the three cultivars are grown in screenhouse conditions. The screenhouse is divided in half and one half of the screenhouse is infested with soybean looper and the other half infested with velvetbean caterpillar. A single leaf is taken from each of the representative plants at 3 different time points, 11 days after infestation, 2 weeks after infestation and 5 weeks after infestation and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation. Total RNA and poly A+ RNA is isolated from pooled tissue consisting of equal quantities of all 18 samples (3 genotypes×3 sample times×2 insect genotypes).

The SOYMON037 cDNA library is generated from soybean cultivar A3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) etiolated axis and radical tissue. Seeds are planted in moist vermiculite, wrapped and kept at room temperature in complete darkness until harvest. Etiolated axis and hypocotyl tissue is harvested at 2, 3 and 4 days post-planting. A total of 1 gram of each tissue type is harvested at 2, 3 and 4 days after planting and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON038 cDNA library is generated from soybean variety Asgrow A3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dry seeds. Explants are prepared for transformation after germination of surface-sterilized seeds on solid tissue media. After 6 days, at 28° C. and 18 hours of light per day, the germinated seeds are cold shocked at 4° C. for 24 hours. Meristemic tissue and part of the hypocotyl is remove and cotyledon excised. The prepared explant is then wounded for Agrobacterium infection. The 2 grams of harvested tissue is frozen in liquid nitrogen and stored at −80° C. until RNA preparation.

The Soy51 (LIB3027) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M280 streptavidin (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The Soy52 (LIB3028) cDNA library is generated from normalized flower DNA. Single stranded DNA representing approximately 1×10⁶ colony forming units of SOYMON022 harvested tissue is used as the starting material for normalization. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M280 streptavidin (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The Soy53 (LIB3039) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling shoot apical meristem tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Apical tissue is harvested from seedling shoot meristem tissue, 7-8 days after the start of imbibition. The apex of each seedling is dissected to include the fifth node to the apical meristem. The fifth node corresponds to the third trifoliate leaf in the very early stages of development. Stipules completely envelop the leaf primordia at this time. A total of 200 mg of apical tissue is harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The Soy54 (LIB3040) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heart to torpedo stage embryo tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are collected and embryos removed from surrounding endosperm and maternal tissues. Embryos from globular to young torpedo stages (by corresponding analogy to Arabidopsis) are collected with a bias towards the middle of this spectrum. Embryos which are beginning to show asymmetric development of cotyledons are considered the upper developmental boundary for the collection and are excluded. A total of 12 mg embryo tissue is frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

Soy55 (LIB3049) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) young seed tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are collected from very young pods (5 to 15 days after flowering). A total of 100 mg of seeds are harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

Soy56 (LIB3029) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M230 streptavidin (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are not converted to double stranded form and represent a non-normalized seed pool for comparison to Soy51 cDNA libraries.

The Soy58 (LIB3050) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa, U.S.A.) drought stressed root tissue subtracted from control root tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr night time cycles. The daytime temperature is approximately 29° and the night time temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days root tissue from both drought stressed and control (watered regularly) plants are collected and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif., U.S.A.). Driver first strand cDNA is covalently linked to DYNADEADS following a protocol similar to that described in the Dynal literature (DYNABEADS. Dynal Corporation. Lake Success, N.Y., U.S.A.). The target cDNA is then heat denatured and the second strand trapped using DYNABEADS oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 4.00 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with DYNABEADS oligo-dT. The tapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif., U.S.A.).

The Soy59 (LIB3051) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) endosperm tissue. Seeds are germinated on paper towels under laboratory ambient light conditions. At 8, 10 and 14 hours after imbibition, the seed coats are harvested. The endosperm consists of a very thin layer of tissue affixed to the inside of the seed coat. The seed coat and endosperm are frozen immediately after harvest in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The Soy60 (LIB3072) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa, U.S.A.) drought stressed seed plus pod subtracted from control seed plus pod tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr night time cycles. The daytime temperature is approximately 26° C. and the night time temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif., U.S.A.). Driver first stand cDNA is covalently linked to DYNABEADS following a protocol similar to that described in the Dynal literature (DYNABEADS Dynal Corporation, Lake Success, N.Y., U.S.A.). The target cDNA is then heat denatured and the second strand trapped using DYNABEADS oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with DYNABEADS oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSEORT vector (Invitrogen, Carlsbad, Calif., U.S.A.).

The Soy61 (LIB3073) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa, U.S.A.) jasmonic acid treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the night time temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo., U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample time points were combined and ground. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories. Palo Alto, Calif., U.S.A.). Driver first strand cDNA is covalently linked to DYNABEADS following a protocol similar to that described in the Dynat literature (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The target cDNA is then heat denatured and the second strand trapped using DYNABEADS oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with DYNABEADS oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif., U.S.A.). For this library's construction, the eighth fraction of the cDNA size fractionation step was used for ligation.

The Soy62 (LIB3074) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa, U.S.A.) jasmonic acid treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the night time temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis. Mo., U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stein is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample time points were combined and ground. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif., U.S.A.). Driver first strand cDNA is covalently linked to DYNABEADS following a protocol similar to that described in the Dynal literature (DYNABEADS, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using DYNABEADS oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with DYNABEADS oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif., U.S.A.). For this library's construction, the ninth fraction of the cDNA size fractionation step was used for ligation.

The Soy65 (LIB3107) 07cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought-stressed abscission zone tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Plants are irrigated with 15-16-17 Peter's Mix. At the R3 stage of development, drought is imposed by withholding water. At 3, 4, 5 and 6 days, tissue is harvested and wilting is not obvious until the fourth day. Abscission layers from reproductive organs are harvested by cutting less than one millimeter proximal and distal to the layer and immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The Soy66 (LIB3109) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) non-drought stressed abscission zone tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Plants are irrigated with 15-16-17 Peter's Mix. At 3, 4, 5 and 6 days, control abscission layer tissue is harvested. Abscission layers from reproductive organs are harvested by cutting less than one millimeter proximal and distal to the layer and immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

Soy67 (LIB3065) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M280 streptavidin (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. Captured hybrids are eluted with water.

Soy68 (LIB3052) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M230 streptavidin (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. Captured hybrids are eluted with water.

Soy69 (LIB3053) cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill., U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) normalized leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr night time cycles. The daytime temperature is approximately 29° C. and the night time temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the C node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on DYNABEADS M280 streptavidin (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are convened to double stranded form and represent the primary normalized library.

Soy70 (LIB3055) cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4^(th) node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

Soy71 (LIB3056) cDNA library is generated from soybean cultivars Cristalina and FT108 (tropical germ plasma) root tissue. Roots are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 50 g and 56 g of roots are harvested from each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

Soy72 (LIB3093) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa, U.S.A.) drought stressed leaf control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr night time cycles. The daytime temperature is approximately 26° C. and the night time temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonerech Laboratories, Palo Alto, Calif., U.S.A.). Driver first strand cDNA is covalently linked to DYNABEADS following a protocol similar to that described in the Dynal literature (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The target cDNA is then heat denatured and the second strand trapped using DYNABEADS oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with DYNABEADS oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif., U.S.A.).

Soy73 (LIB3093) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa, U.S.A.) drought stressed leaf subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr night time cycles. The daytime temperature is approximately 26° C. and the night time temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif., U.S.A.). Driver first strand cDNA is covalently linked to DYNABEADS following a protocol similar to that described in the Dynal literature (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The target cDNA is then heat denatured and the second strand trapped using DYNABEADS oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with DYNABEADS oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Irivitrogen, Carlsbad, Calif., U.S.A.).

The Soy76 (LIB3106) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa, U.S.A.) jasmonic acid and arachidonic treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the night time temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo., U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stein is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. Arachidonic treated seedlings are sprayed with 1 m/1 ml arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample time points were combined and ground. The RNA from the arachidonic treated seedlings is isolated separately. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif., U.S.A.). Driver first strand cDNA is covalently linked to DYNABEADS following a protocol similar to that described in the Dynal literature (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.). The target cDNA is then heat denatured and the second strand trapped using DYNABEADS oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with DYNABEADS oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif., U.S.A.). Fraction 10 of the size fractionated cDNA is ligated into the pSPGRT vector (Invitrogen, Carlsbad, Calif., U.S.A.) in order to capture some of the smaller transcripts characteristic of antifungal proteins.

Soy77 (LIB3108) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the night time temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo., U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid the soybean plantlets appear growth retarded. Arachidonic treated seedlings are sprayed with 1 m/1 ml arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample time points were combined and ground. The RNA from the arachidonic treated seedlings is isolated separately. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif., U.S.A.), Driver first strand cDNA is covalently linked to DYNABEADS following a protocol similar to that described in the Dynal literature (DYNABEADS, Dynal Corporation. Lake Success, N.Y., U.S.A.). The target cDNA is then heat denatured and the second strand trapped using DYNABEADS oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with DYNABEADS oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif., U.S.A.). Fraction 10 of the size fractionated cDNA is ligated into the pSPORT vector in order to capture some of the smaller transcripts characteristic of antifungal proteins.

The stored RNA is purified using Trizol reagent from Life Technologies (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.), essentially as recommended by the manufacturer, Poly A+ RNA (mRNA) is purified using magnetic oligo dT beads essentially as recommended by the manufacturer (DYNABEADS, Dynal Corporation, Lake Success, N.Y., U.S.A.).

Construction of plant cDNA libraries is well-known in the art and a number of cloning strategies exist. A number of cDNA library construction kits are commercially available. The SUPERSCRIPT Plasmid System for cDNA synthesis and Plasmid Cloning (Gibco BRL, Life Technologies, Caithersburg Md., U.S.A.) is used, following the conditions suggested by the manufacturer.

Normalized libraries are made using essentially the Soares procedure (Soares et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:9228-9232 (1994), the entirety of which is herein incorporated by reference). This approach is designed to reduce the initial 10,000-fold variation in individual cDNA frequencies to achieve abundances within one order of magnitude while maintaining the overall sequence complexity of the library. In the normalization process, the prevalence of high-abundance cDNA clones decreases dramatically, clones with mid-level abundance are relatively unaffected and clones for rare transcripts are effectively increased in abundance.

Example 2

The cDNA libraries are plated on LB agar containing the appropriate antibiotics for selection and incubated at 37° for a sufficient time to allow the growth of individual colonies. Single colonies are individually placed in each well of a 96-well microtiter plates containing LB liquid including the selective antibiotics. The plates are incubated overnight at approximately 37° C. with gentle shaking to promote growth of the cultures. The plasmid DNA is isolated from each clone using Qiaprep plasmid isolation kits, using the conditions recommended by the manufacturer (Qiagen Inc., Santa Clara, Calif. U.S.A.).

Template plasmid DNA clones are used for subsequent sequencing. For sequencing, the ABI PRISM dRhodamine Terminator Cycle Sequencing Ready Reaction Kit with AmpliTaq® DNA Polymerase, FS, is used (PE Applied Biosystems, Foster City, Calif. U.S.A.).

Example 3

Nucleic acid sequences that encode for the following proteins: methionine adenosyltransferase, S-adenosylmethionine decarboxylase, aspartate kinase, aspartate-semialdehyde dehydrogenase, O-succinylhomoserine (thiol)-lyase, cystathionine β-lyase, 5-methyltetrahydropteroyltriglutamate, adenosylhomocysteinase, cystathionine β-synthase, cystathionine γ-lyase and O-acetylhomoserine (thiol)-lyase are identified from the Monsanto EST PhytoSeq database using TBLASTN (default values) (TBLASTN compares a protein query against the six reading frames of a nucleic acid sequence). Matches found with BLAST P values equal or less than 0.001 (probability) or BLAST Score of equal or greater than 90 are classified as hits. If the program used to determine the hit is HMMSW then the score refers to HMMSW score.

In addition, the GENBANK database is searched with BLASTN and BLASTX (default values) using ESTs as queries. EST that pass the hit probability threshold of 10e⁻⁸ for the following enzymes are combined with the hits generated by using TBLASTN (described above) and classified by enzyme (see Table A below).

A cluster refers to a set of overlapping clones in the PhytoSeq database. Such an overlapping relationship among clones is designated as a “cluster” when BLAST scores from pairwise sequence comparisons of the member clones meets a predetermined minimum value or product score of 50 or more (Product Score=(BLAST SCORE×Percentage Identity)/(5× minimum [length (Seq1), length (Seq2)]))

Since clusters are formed on the basis of single-linkage relationships, it is possible for two non-overlapping clones to be members of the same cluster if, for instance, they both overlap a third clone with at least the predetermined minimum BLAST score (stringency). A cluster ID is arbitrarily assigned to all of those clones which belong to the same cluster at a given stringency and a particular clone will belong to only one cluster at a given stringency. If a cluster contains only a single clone (a “singleton”), then the cluster ID number will be negative, with an absolute value equal to the clone ID number of its single member. Clones grouped in a cluster in most cases represent a contiguous sequence.

TABLE A* METHIONINE ADENOSYLTRANSFERASE (EC 2.5.1.6) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 1 -700019427 700019427H1 SATMON001 g17262 BLASTX 120 1e−11 92 2 -700074004 700074004H1 SATMON007 g1778820 BLASTN 830 1e−60 80 3 -700149718 700149718H1 SATMON007 g1778820 BLASTN 263 1e−13 80 4 -700220251 700220251H1 SATMON011 g1127582 BLASTN 243 1e−9 90 5 -700458196 700458196H1 SATMON029 g1778820 BLASTN 202 1e−22 88 6 -701172459 701172459H1 SATMONN05 g882334 BLASTN 801 1e−57 89 7 -L1436317 LIB143-062-Q1-E1-B6 LIB143 g1778820 BLASTN 341 1e−19 77 8 -L1893126 LIB189-021-Q1-E1-F4 LIB189 g1778820 BLASTN 629 1e−41 88 9 -L1894542 LIB189-033-Q1-E1-H12 LIB189 g1778820 BLASTN 564 1e−60 76 10 -L30622839 LIB3062-028-Q1-K1-C2 LIB3062 g1778820 BLASTN 625 1e−65 74 11 -L30671966 LIB3067-036-Q1-K1-D8 LIB3067 g1655576 BLASTX 118 1e−25 85 12 -L30682166 LIB3068-004-Q1-K1-D5 LIB3068 g497900 BLASTX 111 1e−29 47 13 1 LIB143-036-Q1-E1-G7 LIB143 g450548 BLASTN 1766 1e−138 88 14 1 LIB3060-013-Q1-K1-F7 LIB3060 g1778820 BLASTN 1702 1e−133 90 15 1 LIB143-013-Q1-E1-A6 LIB143 g1778820 BLASTN 1712 1e−133 88 16 1 LIB148-007-Q1-E1-B8 LIB148 g450548 BLASTN 1693 1e−132 88 17 1 LIB3079-003-Q1-K1-G1 LIB3079 g1778820 BLASTN 1236 1e−130 88 18 1 LIB3066-011-Q1-K1-E4 LIB3066 g1778820 BLASTN 1661 1e−129 86 19 1 LIB3068-008-Q1-K1-D2 LIB3068 g450548 BLASTN 1192 1e−128 88 20 1 LIB143-061-Q1-E1-E7 LIB143 g1778820 BLASTN 1638 1e−127 88 21 1 LIB189-006-Q1-E1-B2 LIB189 g1778820 BLASTN 1627 1e−126 86 22 1 LIB143-031-Q1-E1-C7 LIB143 g450548 BLASTN 1595 1e−124 86 23 1 LIB3068-011-Q1-K1-B12 LIB3068 g450548 BLASTN 1552 1e−123 87 24 1 LIB143-013-Q1-E1-G11 LIB143 g450548 BLASTN 1585 1e−123 88 25 1 LIB3062-057-Q1-K1-H9 LIB3062 g450548 BLASTN 1577 1e−122 89 26 1 LIB143-008-Q1-E1-G9 LIB143 g450548 BLASTN 1556 1e−120 89 27 1 LIB3068-058-Q1-K1-B7 LIB3068 g450548 BLASTN 1470 1e−118 86 28 1 LIB3067-048-Q1-K1-G5 LIB3067 g1778820 BLASTN 1518 1e−117 89 29 1 LIB143-049-Q1-E1-A12 LIB143 g1778820 BLASTN 1480 1e−114 91 30 1 LIB3062-010-Q1-K1-C7 LIB3062 g1778820 BLASTN 1145 1e−113 87 31 1 LIB3068-055-Q1-K1-E10 LIB3068 g450548 BLASTN 1451 1e−112 88 32 1 LIB189-025-Q1-E1-H1 LIB189 g1778820 BLASTN 1460 1e−112 90 33 1 LIB143-066-Q1-E1-F4 LIB143 g1778820 BLASTN 1309 1e−111 86 34 1 LIB143-003-Q1-E1-D10 LIB143 g1778820 BLASTN 1440 1e−111 87 35 1 LIB3062-033-Q1-K1-G1 LIB3062 g1778820 BLASTN 1449 1e−111 86 36 1 LIB3066-029-Q1-K1-H11 LIB3066 g1778820 BLASTN 1427 1e−110 88 37 1 LIB3068-016-Q1-K1-D9 LIB3068 g450548 BLASTN 1213 1e−109 87 38 1 LIB3062-027-Q1-K1-B11 LIB3062 g1778820 BLASTN 1187 1e−108 87 39 1 LIB3068-048-Q1-K1-G4 LIB3068 g1778820 BLASTN 1410 1e−108 82 40 1 LIB148-020-Q1-E1-B6 LIB148 g1778820 BLASTN 1088 1e−107 84 41 1 LIB3066-046-Q1-K1-F2 LIB3066 g1778820 BLASTN 1400 1e−107 90 42 1 700084130H1 SATMON011 g1778820 BLASTN 960 1e−106 92 43 1 700092863H1 SATMON008 g1778820 BLASTN 1388 1e−106 92 44 1 LIB3068-043-Q1-K1-A12 LIB3068 g450548 BLASTN 827 1e−105 81 45 1 700092659H1 SATMON008 g1778820 BLASTN 1359 1e−104 92 46 1 LIB143-038-Q1-E1-H11 LIB143 g450548 BLASTN 1345 1e−103 84 47 1 LIB3062-028-Q1-K1-F12 LIB3062 g1778820 BLASTN 1082 1e−102 82 48 1 700103135H1 SATMON010 g1778820 BLASTN 1306 1e−100 93 49 1 LIB3078-050-Q1-K1-D9 LIB3078 g1778820 BLASTN 1309 1e−100 86 50 1 700084823H1 SATMON011 g1778820 BLASTN 1311 1e−100 90 51 1 700201311H1 SATMON003 g1778820 BLASTN 960 1e−97 89 52 1 700265914H1 SATMON017 g1778820 BLASTN 1275 1e−97 91 53 1 700205948H1 SATMON003 g1778820 BLASTN 983 1e−96 93 54 1 LIB189-003-Q1-E1-A6 LIB189 g450548 BLASTN 1262 1e−96 86 55 1 LIB143-017-Q1-E1-B9 LIB143 g1778820 BLASTN 779 1e−95 86 56 1 700097434H1 SATMON009 g450548 BLASTN 1247 1e−95 90 57 1 700089089H1 SATMON011 g450548 BLASTN 1251 1e−95 89 58 1 700071949H1 SATMON007 g1778820 BLASTN 1256 1e−95 87 59 1 700047892H1 SATMON003 g1778820 BLASTN 1236 1e−94 91 60 1 700077246H1 SATMON007 g450548 BLASTN 1237 1e−94 89 61 1 700085681H1 SATMON011 g1778820 BLASTN 1237 1e−94 89 62 1 LIB3067-010-Q1-K1-A7 LIB3067 g1778820 BLASTN 1244 1e−94 83 63 1 700619961H1 SATMON034 g450548 BLASTN 962 1e−93 89 64 1 700087395H1 SATMON011 g450548 BLASTN 1040 1e−93 89 65 1 700240431H1 SATMON010 g1778820 BLASTN 1156 1e−93 92 66 1 700053822H1 SATMON011 g1778820 BLASTN 1224 1e−93 91 67 1 700103856H1 SATMON010 g450548 BLASTN 1224 1e−93 88 68 1 700083703H1 SATMON011 g450548 BLASTN 1229 1e−93 89 69 1 700087360H1 SATMON011 g450548 BLASTN 1234 1e−93 90 70 1 700104713H1 SATMON010 g450548 BLASTN 1188 1e−92 87 71 1 700265996H1 SATMON017 g450548 BLASTN 1214 1e−92 88 72 1 700264870H1 SATMON017 g450548 BLASTN 945 1e−91 89 73 1 700219348H1 SATMON011 g1778820 BLASTN 1203 1e−91 91 74 1 700095527H1 SATMON008 g450548 BLASTN 1153 1e−90 88 75 1 LIB3068-023-Q1-K1-G5 LIB3068 g1778820 BLASTN 1187 1e−90 89 76 1 700102547H1 SATMON010 g1778820 BLASTN 1194 1e−90 88 77 1 700332179H1 SATMON019 g450548 BLASTN 1054 1e−89 88 78 1 700405470H1 SATMON029 g450548 BLASTN 1071 1e−89 90 79 1 LIB3069-043-Q1-K1-G2 LIB3069 g450548 BLASTN 1175 1e−89 84 80 1 700451332H1 SATMON028 g450548 BLASTN 1176 1e−89 90 81 1 700028108H1 SATMON003 g450548 BLASTN 1180 1e−89 89 82 1 700076154H1 SATMON007 g1778820 BLASTN 1180 1e−89 89 83 1 700089770H1 SATMON011 g450548 BLASTN 1185 1e−89 89 84 1 700051882H1 SATMON003 g1778820 BLASTN 844 1e−88 91 85 1 700242942H1 SATMON010 g1778820 BLASTN 1163 1e−88 91 86 1 700094996H1 SATMON008 g1778820 BLASTN 1166 1e−88 88 87 1 700476239H1 SATMON025 g450548 BLASTN 1166 1e−88 89 88 1 700219530H1 SATMON011 g1778820 BLASTN 1005 1e−87 91 89 1 700049726H1 SATMON003 g1778820 BLASTN 1041 1e−87 89 90 1 700096904H1 SATMON008 g1778820 BLASTN 1157 1e−87 86 91 1 700344026H1 SATMON021 g450548 BLASTN 1158 1e−87 88 92 1 700105590H1 SATMON010 g1778820 BLASTN 1139 1e−86 89 93 1 700465283H1 SATMON025 g450548 BLASTN 1145 1e−86 89 94 1 700104341H1 SATMON010 g450548 BLASTN 1146 1e−86 87 95 1 LIB3062-021-Q1-K1-F12 LIB3062 g1778820 BLASTN 923 1e−85 78 96 1 700105246H1 SATMON010 g450548 BLASTN 930 1e−85 89 97 1 700072146H1 SATMON007 g1778820 BLASTN 1131 1e−85 91 98 1 700028553H1 SATMON003 g1778820 BLASTN 1132 1e−85 88 99 1 700451419H1 SATMON028 g450548 BLASTN 1133 1e−85 89 100 1 700103241H1 SATMON010 g450548 BLASTN 1134 1e−85 90 101 1 700612549H1 SATMON033 g450548 BLASTN 1134 1e−85 89 102 1 700050434H1 SATMON003 g450548 BLASTN 723 1e−84 89 103 1 700094764H1 SATMON008 g1778820 BLASTN 977 1e−84 86 104 1 700163030H1 SATMON013 g1778820 BLASTN 1011 1e−84 93 105 1 700466542H1 SATMON025 g450548 BLASTN 1085 1e−84 89 106 1 700047380H1 SATMON003 g450548 BLASTN 1096 1e−84 89 107 1 700456429H1 SATMON029 g450548 BLASTN 1118 1e−84 88 108 1 700096049H1 SATMON008 g1778820 BLASTN 1122 1e−84 86 109 1 700075906H1 SATMON007 g1778820 BLASTN 838 1e−83 86 110 1 LIB3067-048-Q1-K1-G3 LIB3067 g1778820 BLASTN 1045 1e−83 86 111 1 700214043H1 SATMON016 g450548 BLASTN 1103 1e−83 88 112 1 700096909H1 SATMON008 g1778820 BLASTN 1103 1e−83 90 113 1 701184635H1 SATMONN06 g1778820 BLASTN 1104 1e−83 87 114 1 700158969H1 SATMON012 g1778820 BLASTN 1107 1e−83 93 115 1 700475902H1 SATMON025 g450548 BLASTN 1112 1e−83 89 116 1 700207076H1 SATMON003 g1778820 BLASTN 1113 1e−83 90 117 1 700161901H1 SATMON012 g1778820 BLASTN 1091 1e−82 91 118 1 700452183H1 SATMON028 g450548 BLASTN 1095 1e−82 88 119 1 700452326H1 SATMON028 g450548 BLASTN 1098 1e−82 88 120 1 700106918H1 SATMON010 g450548 BLASTN 1101 1e−82 88 121 1 700220146H1 SATMON011 g450548 BLASTN 1083 1e−81 89 122 1 700221060H1 SATMON011 g450548 BLASTN 1084 1e−81 88 123 1 700243344H1 SATMON010 g1778820 BLASTN 1084 1e−81 92 124 1 700475790H1 SATMON025 g450548 BLASTN 1084 1e−81 89 125 1 700452691H1 SATMON028 g1778820 BLASTN 1086 1e−81 87 126 1 700243733H1 SATMON010 g450548 BLASTN 1086 1e−81 90 127 1 LIB3059-018-Q1-K1-D10 LIB3059 g1778820 BLASTN 830 1e−80 91 128 1 700165958H1 SATMON013 g450548 BLASTN 1070 1e−80 89 129 1 700082659H1 SATMON011 g1778820 BLASTN 1071 1e−80 88 130 1 700172557H1 SATMON013 g1778820 BLASTN 1074 1e−80 92 131 1 700221526H1 SATMON011 g450548 BLASTN 1075 1e−80 88 132 1 700456696H1 SATMON029 g450548 BLASTN 1077 1e−80 89 133 1 700430776H1 SATMONN01 g1778820 BLASTN 574 1e−79 90 134 1 LIB3062-025-Q1-K1-B11 LIB3062 g450548 BLASTN 585 1e−79 85 135 1 700077126H1 SATMON007 g1778820 BLASTN 743 1e−79 88 136 1 700160310H1 SATMON012 g1778820 BLASTN 752 1e−79 91 137 1 700030466H1 SATMON003 g1778820 BLASTN 757 1e−79 83 138 1 700210204H1 SATMON016 g1778820 BLASTN 844 1e−79 91 139 1 700094716H1 SATMON008 g1778820 BLASTN 857 1e−79 86 140 1 700157051H1 SATMON012 g1778820 BLASTN 937 1e−79 89 141 1 700241415H1 SATMON010 g450548 BLASTN 1055 1e−79 90 142 1 700160127H1 SATMON012 g450548 BLASTN 1056 1e−79 88 143 1 700205485H1 SATMON003 g1778820 BLASTN 1059 1e−79 90 144 1 700475930H1 SATMON025 g450548 BLASTN 1060 1e−79 89 145 1 700161902H1 SATMON012 g1778820 BLASTN 1060 1e−79 91 146 1 700213573H1 SATMON016 g450548 BLASTN 1061 1e−79 89 147 1 700207677H1 SATMON016 g450548 BLASTN 1061 1e−79 88 148 1 700581117H1 SATMON031 g450548 BLASTN 1065 1e−79 88 149 1 700451418H1 SATMON028 g450548 BLASTN 546 1e−78 89 150 1 700212606H1 SATMON016 g1778820 BLASTN 1044 1e−78 85 151 1 700222605H1 SATMON011 g1778820 BLASTN 1047 1e−78 86 152 1 700050542H1 SATMON003 g1778820 BLASTN 1053 1e−78 86 153 1 700450709H1 SATMON028 g450548 BLASTN 834 1e−77 88 154 1 700213368H1 SATMON016 g450548 BLASTN 861 1e−77 89 155 1 700095926H1 SATMON008 g1778820 BLASTN 1030 1e−77 86 156 1 700261686H1 SATMON017 g1778820 BLASTN 1034 1e−77 89 157 1 700235993H1 SATMON010 g450548 BLASTN 1037 1e−77 88 158 1 700466152H1 SATMON025 g450548 BLASTN 1038 1e−77 89 159 1 700266410H1 SATMON017 g1778820 BLASTN 1041 1e−77 85 160 1 700216991H1 SATMON016 g1778820 BLASTN 491 1e−76 92 161 1 700464957H1 SATMON025 g450548 BLASTN 607 1e−76 88 162 1 700160463H1 SATMON012 g1778820 BLASTN 1018 1e−76 91 163 1 700158224H1 SATMON012 g1778820 BLASTN 1018 1e−76 92 164 1 700469971H1 SATMON025 g1778820 BLASTN 1020 1e−76 86 165 1 700222731H1 SATMON011 g450548 BLASTN 1022 1e−76 88 166 1 700087434H1 SATMON011 g1778820 BLASTN 1023 1e−76 86 167 1 700094482H1 SATMON008 g1778820 BLASTN 1023 1e−76 86 168 1 700235976H1 SATMON010 g450548 BLASTN 1028 1e−76 89 169 1 700088266H1 SATMON011 g1778820 BLASTN 983 1e−75 92 170 1 700075882H1 SATMON007 g1778820 BLASTN 1009 1e−75 83 171 1 700243324H1 SATMON010 g1778820 BLASTN 1012 1e−75 86 172 1 700159625H2 SATMON012 g450548 BLASTN 1015 1e−75 90 173 1 700204422H1 SATMON003 g1778820 BLASTN 1015 1e−75 88 174 1 700048228H1 SATMON003 g1778820 BLASTN 741 1e−74 85 175 1 700072474H1 SATMON007 g1778820 BLASTN 799 1e−74 85 176 1 700477790H1 SATMON025 g450548 BLASTN 960 1e−74 89 177 1 LIB3059-022-Q1-K1-A9 LIB3059 g1778820 BLASTN 995 1e−74 86 178 1 700241633H1 SATMON010 g450548 BLASTN 997 1e−74 89 179 1 700093978H1 SATMON008 g1778820 BLASTN 999 1e−74 89 180 1 700238569H1 SATMON010 g450548 BLASTN 844 1e−73 88 181 1 700455160H1 SATMON029 g450548 BLASTN 849 1e−73 86 182 1 700262745H1 SATMON017 g1778820 BLASTN 984 1e−73 85 183 1 700405029H1 SATMON027 g1778820 BLASTN 990 1e−73 90 184 1 700195230H1 SATMON014 g450548 BLASTN 991 1e−73 88 185 1 700264753H1 SATMON017 g1778820 BLASTN 993 1e−73 90 186 1 700050142H1 SATMON003 g1778820 BLASTN 433 1e−72 86 187 1 700452677H1 SATMON028 g450548 BLASTN 919 1e−72 87 188 1 700168870H1 SATMON013 g1778820 BLASTN 971 1e−72 90 189 1 700023193H1 SATMON003 g450548 BLASTN 972 1e−72 88 190 1 700086582H1 SATMON011 g1778820 BLASTN 980 1e−72 84 191 1 700457710H1 SATMON029 g450548 BLASTN 982 1e−72 87 192 1 700477601H1 SATMON025 g450548 BLASTN 635 1e−71 88 193 1 700377730H1 SATMON019 g1778820 BLASTN 691 1e−71 87 194 1 700169782H1 SATMON013 g1778820 BLASTN 960 1e−71 91 195 1 700461113H1 SATMON033 g1778820 BLASTN 962 1e−71 85 196 1 700258669H1 SATMON017 g1778820 BLASTN 963 1e−71 90 197 1 700454253H1 SATMON029 g1778820 BLASTN 964 1e−71 85 198 1 700092376H1 SATMON008 g1778820 BLASTN 780 1e−70 83 199 1 700242885H1 SATMON010 g960356 BLASTN 946 1e−70 86 200 1 700094333H1 SATMON008 g1778820 BLASTN 947 1e−70 86 201 1 700575124H1 SATMON030 g450548 BLASTN 948 1e−70 82 202 1 700072544H1 SATMON007 g1778820 BLASTN 949 1e−70 86 203 1 LIB143-046-Q1-E1-B5 LIB143 g450548 BLASTN 951 1e−70 87 204 1 700096534H1 SATMON008 g1778820 BLASTN 956 1e−70 85 205 1 700262906H1 SATMON017 g1778820 BLASTN 937 1e−69 87 206 1 700094478H1 SATMON008 g1778820 BLASTN 939 1e−69 90 207 1 700172985H1 SATMON013 g450548 BLASTN 939 1e−69 88 208 1 700097938H1 SATMON009 g1778820 BLASTN 941 1e−69 85 209 1 LIB3069-006-Q1-K1-D5 LIB3069 g450548 BLASTN 943 1e−69 88 210 1 700093125H1 SATMON008 g1778820 BLASTN 945 1e−69 88 211 1 700424156H1 SATMONN01 g450548 BLASTN 752 1e−68 85 212 1 700405486H1 SATMON029 g450548 BLASTN 864 1e−68 91 213 1 LIB189-026-Q1-E1-H12 LIB189 g1778820 BLASTN 879 1e−68 87 214 1 700106172H1 SATMON010 g1778820 BLASTN 925 1e−68 85 215 1 700096890H1 SATMON008 g1778820 BLASTN 926 1e−68 90 216 1 700154404H1 SATMON007 g450548 BLASTN 928 1e−68 89 217 1 700468337H1 SATMON025 g450548 BLASTN 445 1e−67 88 218 1 700158827H1 SATMON012 g1778820 BLASTN 522 1e−67 87 219 1 700241744H1 SATMON010 g1778820 BLASTN 575 1e−67 85 220 1 700624633H1 SATMON034 g960356 BLASTN 644 1e−67 85 221 1 700154564H1 SATMON007 g1778820 BLASTN 735 1e−67 85 222 1 700172424H1 SATMON013 g450548 BLASTN 912 1e−67 90 223 1 700096287H1 SATMON008 g1778820 BLASTN 914 1e−67 86 224 1 700207638H1 SATMON016 g1778820 BLASTN 914 1e−67 86 225 1 700093735H1 SATMON008 g960356 BLASTN 920 1e−67 89 226 1 700159494H1 SATMON012 g1778820 BLASTN 899 1e−66 89 227 1 700236013H1 SATMON010 g1778820 BLASTN 900 1e−66 83 228 1 700220267H1 SATMON011 g1778820 BLASTN 439 1e−65 84 229 1 700477578H1 SATMON025 g450548 BLASTN 576 1e−65 90 230 1 700047743H1 SATMON003 g1778820 BLASTN 601 1e−65 82 231 1 700343041H1 SATMON021 g1778820 BLASTN 633 1e−65 86 232 1 700159886H1 SATMON012 g450548 BLASTN 803 1e−65 85 233 1 700570242H1 SATMON030 g450548 BLASTN 834 1e−65 85 234 1 700021930H1 SATMON001 g1778820 BLASTN 888 1e−65 87 235 1 700454959H1 SATMON029 g450548 BLASTN 890 1e−65 89 236 1 700171336H1 SATMON013 g1778820 BLASTN 890 1e−65 89 237 1 700105361H1 SATMON010 g1778820 BLASTN 806 1e−64 87 238 1 LIB3068-031-Q1-K1-B1 LIB3068 g450548 BLASTN 853 1e−64 89 239 1 700236324H1 SATMON010 g450548 BLASTN 875 1e−64 89 240 1 700150620H1 SATMON007 g450548 BLASTN 880 1e−64 87 241 1 700220648H1 SATMON011 g450548 BLASTN 881 1e−64 90 242 1 700150733H1 SATMON007 g450548 BLASTN 881 1e−64 85 243 1 700157367H1 SATMON012 g1778820 BLASTN 882 1e−64 85 244 1 700259676H1 SATMON017 g1778820 BLASTN 885 1e−64 88 245 1 700616490H1 SATMON033 g450548 BLASTN 886 1e−64 82 246 1 700102511H1 SATMON010 g450548 BLASTN 695 1e−63 89 247 1 700202805H1 SATMON003 g1778820 BLASTN 817 1e−63 92 248 1 700105685H1 SATMON010 g1778820 BLASTN 866 1e−63 84 249 1 700106113H1 SATMON010 g450548 BLASTN 873 1e−63 91 250 1 700444778H1 SATMON027 g1778820 BLASTN 343 1e−62 84 251 1 700103584H1 SATMON010 g450548 BLASTN 521 1e−62 86 252 1 LIB189-008-Q1-E1-D9 LIB189 g1778820 BLASTN 850 1e−62 91 253 1 700155684H1 SATMON007 g1778820 BLASTN 852 1e−62 85 254 1 700261639H1 SATMON017 g1778820 BLASTN 853 1e−62 89 255 1 700158367H1 SATMON012 g450548 BLASTN 856 1e−62 81 256 1 700153242H1 SATMON007 g1778820 BLASTN 859 1e−62 90 257 1 700210738H1 SATMON016 g1778820 BLASTN 859 1e−62 90 258 1 700203008H1 SATMON003 g450548 BLASTN 698 1e−61 86 259 1 700206081H1 SATMON003 g1778820 BLASTN 840 1e−61 92 260 1 700028643H1 SATMON003 g1778820 BLASTN 846 1e−61 85 261 1 700223914H1 SATMON011 g1778820 BLASTN 849 1e−61 84 262 1 700571455H1 SATMON030 g1778820 BLASTN 378 1e−60 88 263 1 700075374H1 SATMON007 g1778820 BLASTN 830 1e−60 85 264 1 700150452H1 SATMON007 g450548 BLASTN 831 1e−60 89 265 1 700261318H1 SATMON017 g1778820 BLASTN 831 1e−60 83 266 1 700616390H1 SATMON033 g450548 BLASTN 835 1e−60 92 267 1 700208718H1 SATMON016 g450548 BLASTN 561 1e−59 89 268 1 700448948H1 SATMON028 g1778820 BLASTN 653 1e−59 83 269 1 LIB3060-035-Q1-K1-H3 LIB3060 g450548 BLASTN 734 1e−59 85 270 1 700049753H1 SATMON003 g450548 BLASTN 769 1e−59 90 271 1 700154489H1 SATMON007 g1778820 BLASTN 814 1e−59 83 272 1 700170783H1 SATMON013 g1778820 BLASTN 816 1e−59 87 273 1 700237571H1 SATMON010 g450548 BLASTN 817 1e−59 89 274 1 700154872H1 SATMON007 g1778820 BLASTN 822 1e−59 85 275 1 700025620H1 SATMON004 g1778820 BLASTN 686 1e−58 84 276 1 700158255H1 SATMON012 g450548 BLASTN 803 1e−58 86 277 1 700159282H1 SATMON012 g450548 BLASTN 805 1e−58 83 278 1 700222171H1 SATMON011 g1778820 BLASTN 807 1e−58 89 279 1 700205003H1 SATMON003 g1778820 BLASTN 441 1e−57 86 280 1 700049209H1 SATMON003 g1778820 BLASTN 443 1e−57 90 281 1 700016430H1 SATMON001 g1778820 BLASTN 492 1e−57 86 282 1 700212936H1 SATMON016 g450548 BLASTN 564 1e−57 86 283 1 700156939H1 SATMON012 g1778820 BLASTN 801 1e−57 84 284 1 700222183H1 SATMON011 g1778820 BLASTN 561 1e−56 83 285 1 700203453H1 SATMON003 g1778820 BLASTN 784 1e−56 90 286 1 700167708H1 SATMON013 g1778820 BLASTN 560 1e−55 83 287 1 700214356H1 SATMON016 g1778820 BLASTN 693 1e−55 87 288 1 700475377H1 SATMON025 g450548 BLASTN 705 1e−55 90 289 1 700029625H1 SATMON003 g450548 BLASTN 712 1e−55 88 290 1 700202091H1 SATMON003 g1778820 BLASTN 774 1e−55 90 291 1 700264594H1 SATMON017 g450548 BLASTN 775 1e−55 89 292 1 700074969H1 SATMON007 g450548 BLASTN 777 1e−55 87 293 1 LIB3068-005-Q1-K1-A9 LIB3068 g1778820 BLASTN 389 1e−54 80 294 1 700207147H1 SATMON017 g1778820 BLASTN 537 1e−54 88 295 1 LIB189-004-Q1-E1-F11 LIB189 g960356 BLASTN 757 1e−54 87 296 1 700171222H1 SATMON013 g1778820 BLASTN 757 1e−54 93 297 1 700239903H1 SATMON010 g1778820 BLASTN 760 1e−54 84 298 1 700048208H1 SATMON003 g1778820 BLASTN 762 1e−54 90 299 1 700264085H1 SATMON017 g450548 BLASTN 764 1e−54 87 300 1 700155761H1 SATMON007 g1778820 BLASTN 579 1e−53 85 301 1 700216516H1 SATMON016 g450548 BLASTN 380 1e−52 89 302 1 LIB3068-044-Q1-K1-F12 LIB3068 g450548 BLASTN 492 1e−52 73 303 1 700344420H1 SATMON021 g450548 BLASTN 534 1e−52 79 304 1 700219767H1 SATMON011 g450548 BLASTN 732 1e−52 88 305 1 700153757H1 SATMON007 g1778820 BLASTN 737 1e−52 89 306 1 700165841H1 SATMON013 g450548 BLASTN 738 1e−52 90 307 1 700381966H1 SATMON023 g1778820 BLASTN 740 1e−52 86 308 1 700170427H1 SATMON013 g450548 BLASTN 687 1e−51 89 309 1 700094344H1 SATMON008 g1778820 BLASTN 725 1e−51 93 310 1 700052248H1 SATMON003 g1778820 BLASTN 726 1e−51 84 311 1 700050628H1 SATMON003 g450548 BLASTN 726 1e−51 89 312 1 700223031H1 SATMON011 g1778820 BLASTN 729 1e−51 89 313 1 700475686H1 SATMON025 g450548 BLASTN 660 1e−50 89 314 1 700170325H1 SATMON013 g2305013 BLASTN 709 1e−50 82 315 1 700221107H1 SATMON011 g450548 BLASTN 698 1e−49 89 316 1 700612589H1 SATMON033 g960356 BLASTN 703 1e−49 89 317 1 700153770H1 SATMON007 g1778820 BLASTN 705 1e−49 88 318 1 LIB3078-006-Q1-K1-E7 LIB3078 g1778820 BLASTN 705 1e−49 79 319 1 700239724H1 SATMON010 g1778820 BLASTN 507 1e−48 78 320 1 700356777H1 SATMON024 g1778820 BLASTN 683 1e−48 86 321 1 700241568H1 SATMON010 g960356 BLASTN 691 1e−48 88 322 1 700051294H1 SATMON003 g1778820 BLASTN 462 1e−47 88 323 1 700343661H1 SATMON021 g450548 BLASTN 507 1e−47 81 324 1 700029419H1 SATMON003 g1778820 BLASTN 547 1e−47 87 325 1 700165936H1 SATMON013 g1778820 BLASTN 671 1e−47 83 326 1 700026152H1 SATMON003 g960356 BLASTN 673 1e−47 89 327 1 700075671H1 SATMON007 g450548 BLASTN 429 1e−46 89 328 1 700156674H1 SATMON012 g1778820 BLASTN 659 1e−46 91 329 1 700094981H1 SATMON008 g1778820 BLASTN 662 1e−46 85 330 1 700092879H1 SATMON008 g1778820 BLASTN 668 1e−46 87 331 1 700240685H1 SATMON010 g1778820 BLASTN 484 1e−45 84 332 1 700150286H1 SATMON007 g1778820 BLASTN 647 1e−45 82 333 1 700104990H1 SATMON010 g450548 BLASTN 652 1e−45 89 334 1 700203829H1 SATMON003 g450548 BLASTN 652 1e−45 87 335 1 700153718H1 SATMON007 g167961 BLASTN 656 1e−45 91 336 1 700050841H1 SATMON003 g450548 BLASTN 571 1e−44 86 337 1 700268037H1 SATMON017 g2305013 BLASTN 636 1e−44 86 338 1 700153630H1 SATMON007 g1778820 BLASTN 637 1e−44 82 339 1 700475317H1 SATMON025 g450548 BLASTN 530 1e−43 87 340 1 701163127H1 SATMONN04 g960356 BLASTN 626 1e−43 88 341 1 700203688H1 SATMON003 g960356 BLASTN 627 1e−43 89 342 1 700049893H1 SATMON003 g1778820 BLASTN 506 1e−42 90 343 1 700449155H1 SATMON028 g1778820 BLASTN 443 1e−41 85 344 1 701183780H1 SATMONN06 g450548 BLASTN 558 1e−41 86 345 1 700242162H1 SATMON010 g1778820 BLASTN 600 1e−41 90 346 1 700466994H1 SATMON025 g450548 BLASTN 604 1e−41 91 347 1 700259823H1 SATMON017 g450548 BLASTN 607 1e−41 83 348 1 700346242H1 SATMON021 g450548 BLASTN 608 1e−41 86 349 1 700156395H1 SATMON007 g1778820 BLASTN 609 1e−41 90 350 1 700236835H1 SATMON010 g1778820 BLASTN 335 1e−40 81 351 1 700172385H1 SATMON013 g1778820 BLASTN 397 1e−40 87 352 1 700210466H1 SATMON016 g1778820 BLASTN 586 1e−40 81 353 1 700257303H1 SATMON017 g1778820 BLASTN 589 1e−40 81 354 1 LIB3067-027-Q1-K1-G1 LIB3067 g1778820 BLASTN 623 1e−40 87 355 1 LIB3066-025-Q1-K1-D1 LIB3066 g450548 BLASTN 524 1e−39 85 356 1 700106853H1 SATMON010 g450548 BLASTN 580 1e−39 86 357 1 700160540H1 SATMON012 g960356 BLASTN 581 1e−39 88 358 1 700157780H1 SATMON012 g960356 BLASTN 581 1e−39 88 359 1 700149801H1 SATMON007 g450548 BLASTN 565 1e−38 86 360 1 700353243H1 SATMON024 g450548 BLASTN 570 1e−38 86 361 1 700166171H1 SATMON013 g1778820 BLASTN 254 1e−37 79 362 1 700142509H1 SATMON012 g960356 BLASTN 556 1e−37 88 363 1 700242131H1 SATMON010 g450548 BLASTN 560 1e−37 85 364 1 700455643H1 SATMON029 g450548 BLASTN 539 1e−36 86 365 1 700150248H1 SATMON007 g1778820 BLASTN 547 1e−36 87 366 1 700208549H1 SATMON016 g450548 BLASTN 559 1e−36 88 367 1 700027193H1 SATMON003 g450548 BLASTN 529 1e−35 86 368 1 700221390H1 SATMON011 g450548 BLASTN 530 1e−35 85 369 1 700455647H1 SATMON029 g450548 BLASTN 531 1e−35 85 370 1 700260103H1 SATMON017 g1778820 BLASTN 531 1e−35 80 371 1 700167344H1 SATMON013 g1778820 BLASTN 536 1e−35 88 372 1 700089913H1 SATMON011 g960356 BLASTN 546 1e−35 88 373 1 700570573H1 SATMON030 g450548 BLASTN 300 1e−34 89 374 1 700169889H1 SATMON013 g1778820 BLASTN 519 1e−34 89 375 1 700262857H1 SATMON017 g1778820 BLASTN 521 1e−34 87 376 1 700142644H2 SATMON013 g1778820 BLASTN 522 1e−34 87 377 1 700224417H1 SATMON011 g450548 BLASTN 522 1e−34 91 378 1 700073882H1 SATMON007 g450548 BLASTN 531 1e−34 87 379 1 700085803H1 SATMON011 g450548 BLASTN 338 1e−33 89 380 1 700162323H1 SATMON012 g1778820 BLASTN 513 1e−33 86 381 1 700468306H1 SATMON025 g450548 BLASTN 519 1e−33 89 382 1 700443224H2 SATMON026 g450548 BLASTN 275 1e−32 83 383 1 LIB3066-054-Q1-K1-E3 LIB3066 g450548 BLASTN 495 1e−32 87 384 1 700211827H1 SATMON016 g1778820 BLASTN 489 1e−31 80 385 1 700048741H1 SATMON003 g450548 BLASTN 500 1e−31 88 386 1 700613620H1 SATMON033 g1778820 BLASTN 385 1e−30 88 387 1 700029203H1 SATMON003 g1778820 BLASTN 461 1e−29 84 388 1 700378431H1 SATMON020 g450548 BLASTN 466 1e−29 89 389 1 700455641H1 SATMON029 g450548 BLASTN 472 1e−29 85 390 1 700447867H1 SATMON027 g960356 BLASTN 473 1e−29 88 391 1 700447511H1 SATMON027 g450548 BLASTN 474 1e−29 88 392 1 700025851H1 SATMON003 g450548 BLASTN 479 1e−29 88 393 1 LIB3066-024-Q1-K1-H4 LIB3066 g1778820 BLASTN 481 1e−29 81 394 1 LIB143-025-Q1-E1-C4 LIB143 g450549 BLASTX 64 1e−28 70 395 1 700242282H1 SATMON010 g1778820 BLASTN 280 1e−28 85 396 1 700087244H1 SATMON011 g450548 BLASTN 464 1e−28 88 397 1 700458127H1 SATMON029 g450548 BLASTN 238 1e−27 82 398 1 700025767H1 SATMON003 g1778820 BLASTN 438 1e−26 86 399 1 700235401H1 SATMON010 g450548 BLASTN 442 1e−26 88 400 1 700029457H1 SATMON003 g450548 BLASTN 446 1e−26 89 401 1 700266174H1 SATMON017 g450548 BLASTN 447 1e−26 89 402 1 700349254H1 SATMON023 g1778820 BLASTN 315 1e−25 85 403 1 LIB3068-041-Q1-K1-H6 LIB3068 g450548 BLASTN 438 1e−25 86 404 1 700092889H1 SATMON008 g1778820 BLASTN 397 1e−24 92 405 1 700049044H1 SATMON003 g1778820 BLASTN 425 1e−24 90 406 1 700202710H1 SATMON003 g960357 BLASTX 114 1e−19 97 407 1 700155117H1 SATMON007 g450548 BLASTN 314 1e−19 90 408 1 700449619H1 SATMON028 g1778820 BLASTN 341 1e−19 89 409 1 700150336H1 SATMON007 g1778820 BLASTN 343 1e−19 87 410 1 700166223H1 SATMON013 g960357 BLASTX 181 1e−18 97 411 1 700153796H1 SATMON007 g2305013 BLASTN 300 1e−16 82 412 1 700053266H1 SATMON008 g450548 BLASTN 292 1e−15 83 413 1 700405227H1 SATMON028 g450548 BLASTN 313 1e−15 90 414 1 700397410H1 SATMONN01 g17262 BLASTX 147 1e−14 96 415 1 700211524H1 SATMON016 g1033190 BLASTX 153 1e−14 100 416 1 700281415H2 SATMON019 g2315140 BLASTX 139 1e−13 89 417 1 700429873H1 SATMONN01 g16961 BLASTX 133 1e−12 94 418 1 700239660H1 SATMON010 g450548 BLASTN 245 1e−12 86 419 1 700213596H1 SATMON016 g450548 BLASTN 258 1e−12 95 420 1 700152367H1 SATMON007 g450548 BLASTN 273 1e−12 88 421 1 700452014H1 SATMON028 g17262 BLASTX 76 1e−11 72 422 1 700357106H1 SATMON024 g1724104 BLASTX 132 1e−11 100 423 1 700468611H1 SATMON025 g450549 BLASTX 93 1e−10 93 424 1 700213526H1 SATMON016 g450549 BLASTX 127 1e−10 96 425 1 700267065H1 SATMON017 g450549 BLASTX 130 1e−10 96 426 1 700152044H1 SATMON007 g450549 BLASTX 88 1e−8 93 427 1 700159090H1 SATMON012 g169665 BLASTX 113 1e−8 81 428 1 700266734H1 SATMON017 g450549 BLASTX 119 1e−8 96 429 1 700405367H1 SATMON029 g1778820 BLASTN 231 1e−8 65 1635 -700555532 700555532H1 SOYMON001 g429103 BLASTN 920 1e−67 84 1636 -700649594 700649594H1 SOYMON003 g609559 BLASTX 186 1e−23 85 1637 -700750590 700750590H1 SOYMON014 g609224 BLASTN 363 1e−40 79 1638 -700755802 700755802H1 SOYMON014 g609224 BLASTN 479 1e−43 74 1639 -700869211 700869211H1 SOYMON016 g169665 BLASTX 146 1e−13 92 1640 -700891960 700891960H1 SOYMON024 g726031 BLASTN 589 1e−56 82 1641 -700900377 700900377H1 SOYMON027 g1655577 BLASTN 235 1e−8 78 1642 -700902427 700902427H1 SOYMON027 g1655576 BLASTX 151 1e−13 76 1643 -700941686 700941686H1 SOYMON024 g497899 BLASTN 442 1e−26 89 1644 -700952418 700952418H1 SOYMON022 g726030 BLASTX 146 1e−17 83 1645 -700979651 700979651H2 SOYMON009 g1655579 BLASTN 1006 1e−75 84 1646 -700982809 700982809H1 SOYMON009 g1655579 BLASTN 945 1e−69 82 1647 -700982867 700982867H1 SOYMON009 g1127582 BLASTN 868 1e−63 78 1648 -701056884 701056884H1 SOYMON032 g609556 BLASTN 726 1e−51 84 1649 -701117318 701117318H1 SOYMON037 g609224 BLASTN 451 1e−37 80 1650 -701118224 701118224H1 SOYMON037 g2305013 BLASTN 285 1e−19 72 1651 -701121264 701121264H1 SOYMON037 g166873 BLASTN 238 1e−8 82 1652 -701122908 701122908H1 SOYMON037 g16508 BLASTN 444 1e−26 83 1653 -701128589 701128589H1 SOYMON037 g16508 BLASTN 539 1e−36 90 1654 -GM12798 LIB3049-039-Q1-E1-F2 LIB3049 g16508 BLASTN 578 1e−37 73 1655 -GM14331 LIB3049-055-Q1-E1-F5 LIB3049 g167961 BLASTN 497 1e−30 62 1656 -GM30881 LIB3050-005-Q1-K1-G1 LIB3050 g1655577 BLASTN 543 1e−34 82 1657 -GM30911 LIB3050-005-Q1-K1-B8 LIB3050 g1655577 BLASTN 387 1e−26 71 1658 -GM33921 LIB3051-028-Q1-K1-A9 LIB3051 g16508 BLASTN 338 1e−32 73 1659 12644 701131794H1 SOYMON038 g429107 BLASTN 877 1e−64 84 1660 12644 701142515H1 SOYMON038 g429107 BLASTN 871 1e−63 84 1661 12644 700888494H1 SOYMON024 g429107 BLASTN 662 1e−46 83 1662 16 LIB3030-009-Q1-B1-C1 LIB3030 g429105 BLASTN 1439 1e−119 84 1663 16 LIB3051-106-Q1-K1-B5 LIB3051 g609224 BLASTN 1516 1e−117 86 1664 16 LIB3050-023-Q1-K1-A12 LIB3050 g1724103 BLASTN 1388 1e−106 83 1665 16 LIB3028-003-Q1-B1-G11 LIB3028 g609224 BLASTN 1313 1e−100 87 1666 16 LIB3054-010-Q1-N1-E2 LIB3054 g609224 BLASTN 920 1e−95 87 1667 16 700651294H1 SOYMON003 g609224 BLASTN 672 1e−94 86 1668 16 LIB3027-007-Q1-B1-G3 LIB3027 g16508 BLASTN 891 1e−93 83 1669 16 LIB3053-013-Q1-N1-H9 LIB3053 g609224 BLASTN 996 1e−93 87 1670 16 LIB3051-061-Q1-K1-C7 LIB3051 g16508 BLASTN 1195 1e−93 86 1671 16 LIB3065-005-Q1-N1-A6 LIB3065 g609224 BLASTN 707 1e−91 78 1672 16 LIB3050-008-Q1-E1-E3 LIB3050 g609224 BLASTN 1102 1e−87 87 1673 16 LIB3051-011-Q1-E1-B2 LIB3051 g16508 BLASTN 1153 1e−87 82 1674 16 LIB3030-010-Q1-B1-F8 LIB3030 g609224 BLASTN 988 1e−86 83 1675 16 700652104H1 SOYMON003 g1655577 BLASTN 990 1e−86 83 1676 16 701205279H1 SOYMON035 g609556 BLASTN 1125 1e−85 86 1677 16 700662183H1 SOYMON005 g609224 BLASTN 678 1e−83 84 1678 16 LIB3051-039-Q1-K1-F5 LIB3051 g169664 BLASTN 1104 1e−83 86 1679 16 700557616H1 SOYMON001 g609556 BLASTN 1111 1e−83 86 1680 16 LIB3030-002-Q1-B1-C6 LIB3030 g16508 BLASTN 1113 1e−83 83 1681 16 700865235H1 SOYMON016 g1724103 BLASTN 1090 1e−82 84 1682 16 700563340H1 SOYMON002 g609224 BLASTN 911 1e−80 86 1683 16 LIB3040-044-Q1-E1-D7 LIB3040 g166873 BLASTN 1052 1e−80 82 1684 16 LIB3040-060-Q1-E1-D9 LIB3040 g609224 BLASTN 1071 1e−80 84 1685 16 LIB3049-001-Q1-E1-F12 LIB3049 g16508 BLASTN 1074 1e−80 84 1686 16 LIB3028-006-Q1-B1-F1 LIB3028 g16508 BLASTN 857 1e−79 83 1687 16 LIB3049-033-Q1-E1-G5 LIB3049 g2315139 BLASTN 933 1e−79 81 1688 16 700978240H1 SOYMON009 g609224 BLASTN 984 1e−79 88 1689 16 700980802H1 SOYMON009 g862999 BLASTN 1060 1e−79 85 1690 16 700755908H1 SOYMON014 g1724103 BLASTN 1062 1e−79 88 1691 16 700562226H1 SOYMON002 g1724103 BLASTN 1065 1e−79 84 1692 16 701119101H1 SOYMON037 g609224 BLASTN 1048 1e−78 86 1693 16 700646291H1 SOYMON012 g1655577 BLASTN 1049 1e−78 86 1694 16 LIB3050-022-Q1-K1-B9 LIB3050 g16508 BLASTN 1049 1e−78 83 1695 16 LIB3028-030-Q1-B1-F8 LIB3028 g16508 BLASTN 1053 1e−78 83 1696 16 701143128H1 SOYMON038 g609556 BLASTN 737 1e−77 86 1697 16 LIB3040-041-Q1-E1-D10 LIB3040 g166873 BLASTN 861 1e−77 81 1698 16 700756363H1 SOYMON014 g609556 BLASTN 1031 1e−77 88 1699 16 700729963H1 SOYMON009 g1724103 BLASTN 1036 1e−77 88 1700 16 701063046H1 SOYMON033 g609224 BLASTN 1037 1e−77 86 1701 16 LIB3030-005-Q1-B1-G2 LIB3030 g609224 BLASTN 1038 1e−77 84 1702 16 700564331H1 SOYMON002 g609556 BLASTN 1039 1e−77 84 1703 16 700562958H1 SOYMON002 g1724103 BLASTN 1040 1e−77 88 1704 16 LIB3039-014-Q1-E1-F7 LIB3039 g16508 BLASTN 569 1e−76 84 1705 16 700753632H1 SOYMON014 g497899 BLASTN 668 1e−76 83 1706 16 LIB3049-048-Q1-E1-F2 LIB3049 g16508 BLASTN 722 1e−76 83 1707 16 700648911H1 SOYMON003 g1655577 BLASTN 983 1e−76 83 1708 16 LIB3040-027-Q1-E1-H3 LIB3040 g166873 BLASTN 1003 1e−76 81 1709 16 700664905H1 SOYMON005 g609556 BLASTN 1020 1e−76 86 1710 16 701133404H1 SOYMON038 g1724103 BLASTN 1023 1e−76 85 1711 16 701208780H1 SOYMON035 g1655577 BLASTN 1025 1e−76 85 1712 16 700902279H1 SOYMON027 g169664 BLASTN 1026 1e−76 88 1713 16 LIB3040-048-Q1-E1-G10 LIB3040 g16508 BLASTN 1030 1e−76 83 1714 16 LIB3040-028-Q1-E1-A4 LIB3040 g16508 BLASTN 979 1e−75 84 1715 16 700807586H1 SOYMON016 g609224 BLASTN 1006 1e−75 86 1716 16 700755486H1 SOYMON014 g609224 BLASTN 1006 1e−75 87 1717 16 700724920H1 SOYMON009 g609224 BLASTN 1009 1e−75 87 1718 16 701007494H2 SOYMON019 g1724103 BLASTN 1013 1e−75 85 1719 16 700568310H1 SOYMON002 g497899 BLASTN 834 1e−74 83 1720 16 701208819H1 SOYMON035 g609224 BLASTN 857 1e−74 87 1721 16 700985084H1 SOYMON009 g609224 BLASTN 872 1e−74 85 1722 16 701013074H1 SOYMON019 g726031 BLASTN 894 1e−74 88 1723 16 700650843H1 SOYMON003 g609224 BLASTN 924 1e−74 86 1724 16 700646037H1 SOYMON011 g609556 BLASTN 930 1e−74 84 1725 16 LIB3040-039-Q1-E1-H9 LIB3040 g166873 BLASTN 973 1e−74 80 1726 16 700847231H1 SOYMON021 g726031 BLASTN 999 1e−74 85 1727 16 700792293H1 SOYMON011 g609224 BLASTN 999 1e−74 86 1728 16 701109644H1 SOYMON036 g609556 BLASTN 1004 1e−74 86 1729 16 701122929H1 SOYMON037 g1724103 BLASTN 459 1e−73 87 1730 16 700661491H1 SOYMON005 g16508 BLASTN 540 1e−73 90 1731 16 701120070H1 SOYMON037 g609224 BLASTN 786 1e−73 87 1732 16 700898895H1 SOYMON027 g609556 BLASTN 820 1e−73 88 1733 16 701096959H1 SOYMON028 g429105 BLASTN 839 1e−73 82 1734 16 700848356H1 SOYMON021 g497899 BLASTN 985 1e−73 85 1735 16 700864876H1 SOYMON016 g1724103 BLASTN 985 1e−73 88 1736 16 700868456H1 SOYMON016 g609224 BLASTN 987 1e−73 86 1737 16 701210488H1 SOYMON035 g1655577 BLASTN 988 1e−73 83 1738 16 700747764H1 SOYMON013 g609224 BLASTN 822 1e−72 87 1739 16 701063311H1 SOYMON033 g1655577 BLASTN 971 1e−72 86 1740 16 700868625H1 SOYMON016 g1724103 BLASTN 973 1e−72 85 1741 16 700992344H1 SOYMON011 g726031 BLASTN 973 1e−72 85 1742 16 701012719H1 SOYMON019 g1724103 BLASTN 973 1e−72 85 1743 16 700676769H1 SOYMON007 g609224 BLASTN 973 1e−72 87 1744 16 700946235H1 SOYMON024 g609224 BLASTN 975 1e−72 85 1745 16 700891218H1 SOYMON024 g1724103 BLASTN 975 1e−72 84 1746 16 700724907H1 SOYMON009 g609556 BLASTN 977 1e−72 84 1747 16 700833549H1 SOYMON019 g1655577 BLASTN 979 1e−72 85 1748 16 700942105H1 SOYMON024 g726031 BLASTN 980 1e−72 84 1749 16 700564290H1 SOYMON002 g726031 BLASTN 530 1e−71 84 1750 16 700891233H1 SOYMON024 g1724103 BLASTN 746 1e−71 86 1751 16 LIB3028-025-Q1-B1-B2 LIB3028 g609224 BLASTN 870 1e−71 85 1752 16 701120896H1 SOYMON037 g429105 BLASTN 959 1e−71 82 1753 16 LIB3051-027-Q1-K1-A9 LIB3051 g609224 BLASTN 962 1e−71 83 1754 16 701050696H1 SOYMON032 g609556 BLASTN 965 1e−71 86 1755 16 700653619H1 SOYMON003 g609224 BLASTN 966 1e−71 86 1756 16 701047994H1 SOYMON032 g1724103 BLASTN 967 1e−71 85 1757 16 701123056H1 SOYMON037 g1724103 BLASTN 968 1e−71 85 1758 16 700983479H1 SOYMON009 g726031 BLASTN 653 1e−70 85 1759 16 701063733H1 SOYMON034 g1655577 BLASTN 850 1e−70 85 1760 16 700738366H1 SOYMON012 g726031 BLASTN 946 1e−70 85 1761 16 700866319H1 SOYMON016 g1655575 BLASTN 947 1e−70 85 1762 16 700789013H2 SOYMON011 g1655577 BLASTN 948 1e−70 85 1763 16 700896080H1 SOYMON027 g609224 BLASTN 949 1e−70 87 1764 16 LIB3039-021-Q1-E1-F12 LIB3039 g16508 BLASTN 950 1e−70 83 1765 16 700898284H1 SOYMON027 g429105 BLASTN 950 1e−70 86 1766 16 700901743H1 SOYMON027 g609224 BLASTN 950 1e−70 86 1767 16 700831048H1 SOYMON019 g429105 BLASTN 951 1e−70 85 1768 16 701038107H1 SOYMON029 g726031 BLASTN 952 1e−70 87 1769 16 700559703H1 SOYMON001 g726031 BLASTN 954 1e−70 88 1770 16 700848817H1 SOYMON021 g609224 BLASTN 954 1e−70 84 1771 16 700896032H1 SOYMON027 g609224 BLASTN 956 1e−70 87 1772 16 700944764H1 SOYMON024 g16508 BLASTN 567 1e−69 85 1773 16 701011015H1 SOYMON019 g1724103 BLASTN 647 1e−69 81 1774 16 701125662H1 SOYMON037 g609224 BLASTN 751 1e−69 87 1775 16 701046895H1 SOYMON032 g16508 BLASTN 795 1e−69 83 1776 16 701012632H1 SOYMON019 g1655577 BLASTN 800 1e−69 85 1777 16 701005244H1 SOYMON019 g1724103 BLASTN 804 1e−69 82 1778 16 LIB3050-023-Q1-K1-H10 LIB3050 g609224 BLASTN 899 1e−69 83 1779 16 700892558H1 SOYMON024 g1655575 BLASTN 936 1e−69 85 1780 16 700988779H1 SOYMON011 g16508 BLASTN 937 1e−69 82 1781 16 701203923H2 SOYMON035 g429105 BLASTN 938 1e−69 86 1782 16 701010231H2 SOYMON019 g1724103 BLASTN 938 1e−69 83 1783 16 701041790H1 SOYMON029 g450548 BLASTN 939 1e−69 84 1784 16 700967887H1 SOYMON033 g1724103 BLASTN 940 1e−69 87 1785 16 701123361H1 SOYMON037 g16508 BLASTN 941 1e−69 84 1786 16 701123154H1 SOYMON037 g1724103 BLASTN 942 1e−69 86 1787 16 701045767H1 SOYMON032 g726031 BLASTN 942 1e−69 85 1788 16 700983288H1 SOYMON009 g609224 BLASTN 945 1e−69 83 1789 16 700653053H1 SOYMON003 g609224 BLASTN 945 1e−69 86 1790 16 701044226H1 SOYMON032 g1655577 BLASTN 486 1e−68 83 1791 16 700969927H1 SOYMON005 g497899 BLASTN 750 1e−68 85 1792 16 700547956H1 SOYMON001 g609224 BLASTN 779 1e−68 87 1793 16 LIB3049-048-Q1-E1-E4 LIB3049 g609224 BLASTN 827 1e−68 83 1794 16 701140780H1 SOYMON038 g1655577 BLASTN 922 1e−68 85 1795 16 700849166H1 SOYMON021 g1724103 BLASTN 923 1e−68 84 1796 16 700891945H1 SOYMON024 g609556 BLASTN 925 1e−68 87 1797 16 700658051H1 SOYMON004 g429105 BLASTN 925 1e−68 83 1798 16 700942415H1 SOYMON024 g1655575 BLASTN 928 1e−68 84 1799 16 701041890H1 SOYMON029 g1724103 BLASTN 930 1e−68 82 1800 16 LIB3028-006-Q1-B1-H12 LIB3028 g16508 BLASTN 931 1e−68 81 1801 16 700967039H1 SOYMON029 g429105 BLASTN 931 1e−68 85 1802 16 700836178H1 SOYMON019 g1724103 BLASTN 933 1e−68 85 1803 16 700897558H1 SOYMON027 g16508 BLASTN 737 1e−67 85 1804 16 LIB3050-004-Q1-E1-A2 LIB3050 g609224 BLASTN 796 1e−67 80 1805 16 700730236H1 SOYMON009 g726031 BLASTN 816 1e−67 84 1806 16 700833936H1 SOYMON019 g1724103 BLASTN 915 1e−67 84 1807 16 700897552H1 SOYMON027 g1655577 BLASTN 916 1e−67 84 1808 16 700945440H1 SOYMON024 g609556 BLASTN 917 1e−67 84 1809 16 700961368H1 SOYMON022 g169664 BLASTN 918 1e−67 88 1810 16 700789702H1 SOYMON011 g609224 BLASTN 921 1e−67 86 1811 16 700786541H1 SOYMON011 g429105 BLASTN 513 1e−66 84 1812 16 700987282H1 SOYMON009 g16508 BLASTN 523 1e−66 87 1813 16 700661002H1 SOYMON005 g2305013 BLASTN 836 1e−66 81 1814 16 701148312H1 SOYMON031 g862999 BLASTN 899 1e−66 84 1815 16 700738822H1 SOYMON012 g1655577 BLASTN 899 1e−66 85 1816 16 700940996H1 SOYMON024 g609556 BLASTN 901 1e−66 86 1817 16 700749195H1 SOYMON013 g609556 BLASTN 903 1e−66 81 1818 16 700893941H1 SOYMON024 g429105 BLASTN 903 1e−66 88 1819 16 700892888H1 SOYMON024 g609556 BLASTN 906 1e−66 86 1820 16 700901481H1 SOYMON027 g169664 BLASTN 638 1e−65 86 1821 16 700945269H1 SOYMON024 g169664 BLASTN 688 1e−65 86 1822 16 700746876H1 SOYMON013 g609556 BLASTN 740 1e−65 85 1823 16 700755043H1 SOYMON014 g609556 BLASTN 760 1e−65 87 1824 16 701097166H1 SOYMON028 g1655577 BLASTN 781 1e−65 82 1825 16 701129484H1 SOYMON037 g16508 BLASTN 898 1e−65 82 1826 16 700651014H1 SOYMON003 g167961 BLASTN 464 1e−64 81 1827 16 701134363H1 SOYMON038 g726031 BLASTN 687 1e−64 84 1828 16 701124677H1 SOYMON037 g726031 BLASTN 876 1e−64 85 1829 16 701000359H1 SOYMON018 g1724103 BLASTN 877 1e−64 87 1830 16 701139375H1 SOYMON038 g1724103 BLASTN 883 1e−64 84 1831 16 700832784H1 SOYMON019 g609224 BLASTN 883 1e−64 89 1832 16 700980227H1 SOYMON009 g1724103 BLASTN 884 1e−64 84 1833 16 700943177H1 SOYMON024 g1655577 BLASTN 885 1e−64 85 1834 16 700844446H1 SOYMON021 g2315139 BLASTN 355 1e−63 84 1835 16 700836453H1 SOYMON020 g862999 BLASTN 494 1e−63 85 1836 16 700983012H1 SOYMON009 g16508 BLASTN 756 1e−63 83 1837 16 700795805H1 SOYMON017 g1655577 BLASTN 833 1e−63 85 1838 16 701213663H1 SOYMON035 g1724103 BLASTN 862 1e−63 84 1839 16 700868366H1 SOYMON016 g167961 BLASTN 864 1e−63 83 1840 16 701134377H1 SOYMON038 g2305013 BLASTN 865 1e−63 79 1841 16 LIB3049-007-Q1-E1-C9 LIB3049 g16508 BLASTN 867 1e−63 79 1842 16 700944577H1 SOYMON024 g862999 BLASTN 869 1e−63 85 1843 16 700992289H1 SOYMON011 g16508 BLASTN 869 1e−63 84 1844 16 700891612H1 SOYMON024 g609556 BLASTN 872 1e−63 86 1845 16 700738277H1 SOYMON012 g609224 BLASTN 463 1e−62 85 1846 16 700895571H1 SOYMON027 g609224 BLASTN 562 1e−62 88 1847 16 LIB3049-008-Q1-E1-B3 LIB3049 g16508 BLASTN 853 1e−62 77 1848 16 LIB3027-010-Q1-B1-E12 LIB3027 g609224 BLASTN 854 1e−62 82 1849 16 701129389H1 SOYMON037 g16508 BLASTN 854 1e−62 83 1850 16 701045542H1 SOYMON032 g429105 BLASTN 855 1e−62 86 1851 16 700969901H1 SOYMON005 g726031 BLASTN 856 1e−62 84 1852 16 700984664H1 SOYMON009 g1724103 BLASTN 856 1e−62 81 1853 16 700561352H1 SOYMON002 g609224 BLASTN 858 1e−62 85 1854 16 701138828H1 SOYMON038 g16508 BLASTN 858 1e−62 84 1855 16 700845962H1 SOYMON021 g1655577 BLASTN 861 1e−62 87 1856 16 700896147H1 SOYMON027 g16508 BLASTN 336 1e−61 82 1857 16 701137929H1 SOYMON038 g1655577 BLASTN 502 1e−61 84 1858 16 LIB3040-032-Q1-E1-B8 LIB3040 g16508 BLASTN 521 1e−61 82 1859 16 701009537H1 SOYMON019 g726031 BLASTN 545 1e−61 86 1860 16 LIB3049-031-Q1-E1-E4 LIB3049 g609224 BLASTN 677 1e−61 82 1861 16 LIB3049-031-Q1-E1-C7 LIB3049 g167961 BLASTN 696 1e−61 82 1862 16 700891843H1 SOYMON024 g1655577 BLASTN 737 1e−61 84 1863 16 700561231H1 SOYMON002 g16508 BLASTN 839 1e−61 82 1864 16 700548238H1 SOYMON002 g429105 BLASTN 843 1e−61 85 1865 16 700901018H1 SOYMON027 g2305013 BLASTN 845 1e−61 82 1866 16 701134413H1 SOYMON038 g2305013 BLASTN 848 1e−61 82 1867 16 700653524H1 SOYMON003 g609224 BLASTN 499 1e−60 85 1868 16 LIB3049-017-Q1-E1-G8 LIB3049 g16508 BLASTN 514 1e−60 82 1869 16 701121077H1 SOYMON037 g16508 BLASTN 671 1e−60 84 1870 16 LIB3056-013-Q1-N1-A9 LIB3056 g609224 BLASTN 784 1e−60 86 1871 16 700943524H1 SOYMON024 g16508 BLASTN 831 1e−60 85 1872 16 700952889H1 SOYMON022 g429103 BLASTN 835 1e−60 84 1873 16 700730632H1 SOYMON009 g1655577 BLASTN 431 1e−59 82 1874 16 700649136H1 SOYMON003 g609224 BLASTN 486 1e−59 84 1875 16 700895591H1 SOYMON027 g609556 BLASTN 492 1e−59 87 1876 16 701009829H1 SOYMON019 g429103 BLASTN 572 1e−59 80 1877 16 LIB3039-011-Q1-E1-C11 LIB3039 g16508 BLASTN 619 1e−59 83 1878 16 LIB3040-055-Q1-E1-C4 LIB3040 g16508 BLASTN 622 1e−59 82 1879 16 701123571H1 SOYMON037 g2305013 BLASTN 682 1e−59 81 1880 16 LIB3049-004-Q1-E1-E5 LIB3049 g167961 BLASTN 771 1e−59 83 1881 16 701002239H1 SOYMON018 g609224 BLASTN 782 1e−59 86 1882 16 700971088H1 SOYMON005 g1724103 BLASTN 793 1e−59 81 1883 16 700864624H1 SOYMON016 g167961 BLASTN 817 1e−59 83 1884 16 700863534H1 SOYMON027 g16508 BLASTN 818 1e−59 86 1885 16 700749489H1 SOYMON013 g16508 BLASTN 823 1e−59 86 1886 16 700730689H1 SOYMON009 g16508 BLASTN 825 1e−59 83 1887 16 LIB3049-050-Q1-E1-A11 LIB3049 g16508 BLASTN 831 1e−59 83 1888 16 700850803H1 SOYMON023 g1655577 BLASTN 511 1e−58 83 1889 16 700992317H1 SOYMON011 g1655577 BLASTN 520 1e−58 79 1890 16 701119129H1 SOYMON037 g16508 BLASTN 553 1e−58 83 1891 16 700657623H1 SOYMON004 g1724103 BLASTN 600 1e−58 83 1892 16 LIB3049-015-Q1-E1-F8 LIB3049 g16508 BLASTN 674 1e−58 79 1893 16 701007027H1 SOYMON019 g16508 BLASTN 804 1e−58 81 1894 16 701120820H1 SOYMON037 g609224 BLASTN 807 1e−58 85 1895 16 700654317H1 SOYMON004 g1655577 BLASTN 811 1e−58 84 1896 16 700889071H1 SOYMON024 g497899 BLASTN 812 1e−58 83 1897 16 701010676H1 SOYMON019 g609224 BLASTN 813 1e−58 85 1898 16 701099590H1 SOYMON028 g1655577 BLASTN 486 1e−57 82 1899 16 700649684H1 SOYMON003 g16508 BLASTN 540 1e−57 82 1900 16 700994107H1 SOYMON011 g16508 BLASTN 567 1e−57 80 1901 16 700898629H1 SOYMON027 g16508 BLASTN 687 1e−57 82 1902 16 700902333H1 SOYMON027 g609224 BLASTN 700 1e−57 82 1903 16 LIB3039-017-Q1-E1-D9 LIB3039 g16508 BLASTN 703 1e−57 81 1904 16 LIB3040-026-Q1-E1-A4 LIB3040 g16508 BLASTN 709 1e−57 83 1905 16 701130101H1 SOYMON037 g1724103 BLASTN 791 1e−57 88 1906 16 700902482H1 SOYMON027 g169664 BLASTN 797 1e−57 81 1907 16 700730789H1 SOYMON009 g16508 BLASTN 798 1e−57 82 1908 16 700868670H1 SOYMON016 g16508 BLASTN 800 1e−57 83 1909 16 LIB3029-011-Q1-B1-D1 LIB3029 g609224 BLASTN 800 1e−57 82 1910 16 701119278H1 SOYMON037 g609224 BLASTN 801 1e−57 83 1911 16 700747731H1 SOYMON013 g16508 BLASTN 801 1e−57 86 1912 16 LIB3049-017-Q1-E1-A11 LIB3049 g16508 BLASTN 811 1e−57 80 1913 16 700890428H1 SOYMON024 g609224 BLASTN 482 1e−56 83 1914 16 700562326H1 SOYMON002 g609224 BLASTN 778 1e−56 82 1915 16 700567740H1 SOYMON002 g609224 BLASTN 781 1e−56 85 1916 16 701061514H1 SOYMON033 g1724103 BLASTN 784 1e−56 86 1917 16 701135670H1 SOYMON038 g429105 BLASTN 785 1e−56 81 1918 16 701139657H1 SOYMON038 g609224 BLASTN 786 1e−56 85 1919 16 701123371H1 SOYMON037 g16508 BLASTN 788 1e−56 86 1920 16 700838744H1 SOYMON020 g16508 BLASTN 789 1e−56 82 1921 16 701070458H1 SOYMON034 g1655577 BLASTN 457 1e−55 78 1922 16 701003437H1 SOYMON019 g429105 BLASTN 474 1e−55 83 1923 16 700835976H1 SOYMON019 g16508 BLASTN 562 1e−55 84 1924 16 700894535H1 SOYMON024 g16508 BLASTN 570 1e−55 86 1925 16 700752755H1 SOYMON014 g1655577 BLASTN 690 1e−55 83 1926 16 LIB3049-032-Q1-E1-C9 LIB3049 g16508 BLASTN 700 1e−55 83 1927 16 LIB3040-030-Q1-E1-B5 LIB3040 g16508 BLASTN 709 1e−55 84 1928 16 701136935H1 SOYMON038 g609224 BLASTN 766 1e−55 85 1929 16 700682088H1 SOYMON008 g169664 BLASTN 767 1e−55 90 1930 16 700682188H1 SOYMON008 g169664 BLASTN 767 1e−55 90 1931 16 701068649H1 SOYMON034 g16508 BLASTN 770 1e−55 83 1932 16 700726623H1 SOYMON009 g16508 BLASTN 771 1e−55 85 1933 16 700751129H1 SOYMON014 g16508 BLASTN 772 1e−55 86 1934 16 700945234H1 SOYMON024 g16508 BLASTN 773 1e−55 83 1935 16 701133507H2 SOYMON038 g609224 BLASTN 776 1e−55 85 1936 16 700902459H1 SOYMON027 g726031 BLASTN 777 1e−55 84 1937 16 700986624H1 SOYMON009 g16508 BLASTN 612 1e−54 83 1938 16 701097020H1 SOYMON028 g609224 BLASTN 615 1e−54 84 1939 16 701131409H1 SOYMON038 g16508 BLASTN 755 1e−54 91 1940 16 700750123H1 SOYMON013 g726031 BLASTN 756 1e−54 86 1941 16 701040251H1 SOYMON029 g16508 BLASTN 756 1e−54 85 1942 16 700732290H1 SOYMON010 g169664 BLASTN 758 1e−54 90 1943 16 701117458H1 SOYMON037 g609224 BLASTN 759 1e−54 82 1944 16 701118709H1 SOYMON037 g16508 BLASTN 760 1e−54 91 1945 16 701012834H1 SOYMON019 g16508 BLASTN 760 1e−54 91 1946 16 701133316H1 SOYMON038 g16508 BLASTN 760 1e−54 91 1947 16 701129646H1 SOYMON037 g16508 BLASTN 760 1e−54 91 1948 16 700732265H1 SOYMON010 g169664 BLASTN 760 1e−54 90 1949 16 701040796H1 SOYMON029 g1724103 BLASTN 760 1e−54 89 1950 16 700846350H1 SOYMON021 g16508 BLASTN 761 1e−54 86 1951 16 701137005H1 SOYMON038 g16508 BLASTN 761 1e−54 86 1952 16 701046506H1 SOYMON032 g16508 BLASTN 761 1e−54 91 1953 16 700894462H1 SOYMON024 g16508 BLASTN 761 1e−54 86 1954 16 700973847H1 SOYMON005 g16508 BLASTN 766 1e−54 87 1955 16 701128215H1 SOYMON037 g609556 BLASTN 466 1e−53 82 1956 16 700842468H1 SOYMON020 g429105 BLASTN 548 1e−53 86 1957 16 701051552H1 SOYMON032 g609224 BLASTN 744 1e−53 85 1958 16 700944049H1 SOYMON024 g609224 BLASTN 744 1e−53 85 1959 16 701120261H1 SOYMON037 g609224 BLASTN 746 1e−53 84 1960 16 700897524H1 SOYMON027 g609224 BLASTN 751 1e−53 84 1961 16 LIB3049-042-Q1-E1-F7 LIB3049 g450548 BLASTN 761 1e−53 75 1962 16 700896909H1 SOYMON027 g429105 BLASTN 536 1e−52 82 1963 16 700974820H1 SOYMON005 g1724103 BLASTN 582 1e−52 86 1964 16 701143224H1 SOYMON038 g609224 BLASTN 731 1e−52 84 1965 16 701010322H1 SOYMON019 g609224 BLASTN 731 1e−52 85 1966 16 701130510H1 SOYMON038 g609224 BLASTN 732 1e−52 83 1967 16 700724904H1 SOYMON009 g609224 BLASTN 733 1e−52 85 1968 16 701119845H1 SOYMON037 g609224 BLASTN 734 1e−52 85 1969 16 701107871H1 SOYMON036 g16508 BLASTN 737 1e−52 86 1970 16 700983380H1 SOYMON009 g609224 BLASTN 738 1e−52 84 1971 16 700896469H1 SOYMON027 g16508 BLASTN 741 1e−52 86 1972 16 700792178H1 SOYMON011 g16508 BLASTN 742 1e−52 85 1973 16 701099906H1 SOYMON028 g16508 BLASTN 431 1e−51 89 1974 16 701134724H2 SOYMON038 g16508 BLASTN 553 1e−51 85 1975 16 700749712H1 SOYMON013 g16508 BLASTN 689 1e−51 87 1976 16 700982567H1 SOYMON009 g609224 BLASTN 723 1e−51 85 1977 16 701013758H1 SOYMON019 g609224 BLASTN 723 1e−51 85 1978 16 700868508H1 SOYMON016 g609224 BLASTN 723 1e−51 85 1979 16 700749316H1 SOYMON013 g609224 BLASTN 723 1e−51 85 1980 16 701045367H1 SOYMON032 g16508 BLASTN 724 1e−51 91 1981 16 701205494H1 SOYMON035 g609224 BLASTN 724 1e−51 84 1982 16 700556784H1 SOYMON001 g16508 BLASTN 724 1e−51 91 1983 16 701099879H1 SOYMON028 g16508 BLASTN 726 1e−51 80 1984 16 701131671H1 SOYMON038 g609224 BLASTN 728 1e−51 85 1985 16 701011505H1 SOYMON019 g609224 BLASTN 728 1e−51 85 1986 16 701009973H2 SOYMON019 g609224 BLASTN 728 1e−51 85 1987 16 700793520H1 SOYMON017 g609224 BLASTN 728 1e−51 85 1988 16 700753622H1 SOYMON014 g609224 BLASTN 728 1e−51 85 1989 16 700984052H1 SOYMON009 g609224 BLASTN 728 1e−51 83 1990 16 701108521H1 SOYMON036 g609224 BLASTN 728 1e−51 85 1991 16 700957203H1 SOYMON022 g1724103 BLASTN 464 1e−50 86 1992 16 700554120H1 SOYMON001 g609224 BLASTN 528 1e−50 84 1993 16 700555954H1 SOYMON001 g167961 BLASTN 528 1e−50 86 1994 16 701124141H1 SOYMON037 g609224 BLASTN 537 1e−50 85 1995 16 701003232H1 SOYMON019 g16508 BLASTN 544 1e−50 88 1996 16 700653112H1 SOYMON003 g16508 BLASTN 559 1e−50 91 1997 16 701103353H1 SOYMON028 g1655577 BLASTN 581 1e−50 84 1998 16 700562790H1 SOYMON002 g497899 BLASTN 708 1e−50 86 1999 16 701097303H1 SOYMON028 g609224 BLASTN 711 1e−50 84 2000 16 700561195H1 SOYMON002 g609224 BLASTN 712 1e−50 84 2001 16 701121162H1 SOYMON037 g16508 BLASTN 712 1e−50 88 2002 16 700981317H1 SOYMON009 g16508 BLASTN 714 1e−50 89 2003 16 700981595H1 SOYMON009 g16508 BLASTN 716 1e−50 86 2004 16 700994305H1 SOYMON011 g609224 BLASTN 528 1e−49 85 2005 16 701101565H1 SOYMON028 g429105 BLASTN 552 1e−49 82 2006 16 701103456H1 SOYMON028 g429105 BLASTN 617 1e−49 81 2007 16 700993331H1 SOYMON011 g16508 BLASTN 622 1e−49 86 2008 16 LIB3052-011-Q1-N1-B12 LIB3052 g16508 BLASTN 695 1e−49 85 2009 16 701140613H1 SOYMON038 g609224 BLASTN 697 1e−49 85 2010 16 700745426H1 SOYMON013 g16508 BLASTN 697 1e−49 84 2011 16 701046530H1 SOYMON032 g609224 BLASTN 697 1e−49 84 2012 16 701003787H1 SOYMON019 g609224 BLASTN 697 1e−49 85 2013 16 700840830H1 SOYMON020 g1724103 BLASTN 699 1e−49 79 2014 16 700901588H1 SOYMON027 g609224 BLASTN 702 1e−49 85 2015 16 701037824H1 SOYMON029 g497899 BLASTN 702 1e−49 87 2016 16 701131888H1 SOYMON038 g609224 BLASTN 702 1e−49 85 2017 16 701009947H2 SOYMON019 g609224 BLASTN 702 1e−49 85 2018 16 700729216H1 SOYMON009 g497899 BLASTN 702 1e−49 87 2019 16 700831705H1 SOYMON019 g609224 BLASTN 702 1e−49 85 2020 16 700900329H1 SOYMON027 g429105 BLASTN 705 1e−49 88 2021 16 700563946H1 SOYMON002 g16508 BLASTN 705 1e−49 83 2022 16 700808370H1 SOYMON024 g609556 BLASTN 459 1e−48 83 2023 16 LIB3040-001-Q1-E1-H4 LIB3040 g1724103 BLASTN 475 1e−48 76 2024 16 700989482H1 SOYMON011 g16508 BLASTN 498 1e−48 86 2025 16 701001311H1 SOYMON018 g16508 BLASTN 655 1e−48 85 2026 16 700734733H1 SOYMON010 g497899 BLASTN 682 1e−48 87 2027 16 701013070H1 SOYMON019 g497899 BLASTN 682 1e−48 87 2028 16 701120989H1 SOYMON037 g16508 BLASTN 683 1e−48 91 2029 16 LIB3049-025-Q1-E1-B4 LIB3049 g167961 BLASTN 687 1e−48 82 2030 16 700808455H1 SOYMON024 g16508 BLASTN 688 1e−48 87 2031 16 701101319H1 SOYMON028 g16508 BLASTN 688 1e−48 91 2032 16 701037087H1 SOYMON029 g609224 BLASTN 689 1e−48 85 2033 16 701205225H1 SOYMON035 g16508 BLASTN 689 1e−48 84 2034 16 701136807H1 SOYMON038 g609224 BLASTN 690 1e−48 85 2035 16 701118459H1 SOYMON037 g609224 BLASTN 690 1e−48 85 2036 16 700942825H1 SOYMON024 g16508 BLASTN 691 1e−48 85 2037 16 701139796H1 SOYMON038 g497899 BLASTN 692 1e−48 87 2038 16 700557040H1 SOYMON001 g609224 BLASTN 692 1e−48 85 2039 16 701015715H1 SOYMON038 g497899 BLASTN 692 1e−48 87 2040 16 700726424H1 SOYMON009 g497899 BLASTN 693 1e−48 87 2041 16 700790811H1 SOYMON011 g497899 BLASTN 693 1e−48 87 2042 16 LIB3040-038-Q1-E1-E5 LIB3040 g16508 BLASTN 709 1e−48 85 2043 16 700896626H1 SOYMON027 g429107 BLASTN 430 1e−47 84 2044 16 700562158H1 SOYMON002 g609224 BLASTN 518 1e−47 84 2045 16 700747095H1 SOYMON013 g16508 BLASTN 538 1e−47 87 2046 16 700726532H1 SOYMON009 g609224 BLASTN 622 1e−47 85 2047 16 701137686H1 SOYMON038 g497899 BLASTN 671 1e−47 85 2048 16 701009148H1 SOYMON019 g16508 BLASTN 673 1e−47 91 2049 16 701048279H1 SOYMON032 g16508 BLASTN 674 1e−47 86 2050 16 701120185H1 SOYMON037 g16508 BLASTN 674 1e−47 91 2051 16 701040355H1 SOYMON029 g497899 BLASTN 675 1e−47 86 2052 16 701130203H1 SOYMON037 g609224 BLASTN 678 1e−47 83 2053 16 701015794H1 SOYMON038 g16508 BLASTN 681 1e−47 91 2054 16 700732181H1 SOYMON010 g16508 BLASTN 681 1e−47 86 2055 16 LIB3050-018-Q1-E1-D10 LIB3050 g2305013 BLASTN 696 1e−47 81 2056 16 700899157H1 SOYMON027 g609224 BLASTN 486 1e−46 84 2057 16 701119905H1 SOYMON037 g497899 BLASTN 507 1e−46 86 2058 16 701062873H1 SOYMON033 g497899 BLASTN 509 1e−46 88 2059 16 701210886H1 SOYMON035 g16508 BLASTN 540 1e−46 84 2060 16 700893278H1 SOYMON024 g16508 BLASTN 542 1e−46 77 2061 16 701036970H1 SOYMON029 g16508 BLASTN 579 1e−46 88 2062 16 700901932H1 SOYMON027 g609224 BLASTN 640 1e−46 85 2063 16 701044214H1 SOYMON032 g429105 BLASTN 658 1e−46 85 2064 16 701056917H1 SOYMON033 g16508 BLASTN 661 1e−46 85 2065 16 700981560H1 SOYMON009 g497899 BLASTN 661 1e−46 87 2066 16 701213107H1 SOYMON035 g609224 BLASTN 661 1e−46 83 2067 16 700834235H1 SOYMON019 g497899 BLASTN 661 1e−46 87 2068 16 700749413H1 SOYMON013 g16508 BLASTN 662 1e−46 85 2069 16 700889148H1 SOYMON024 g16508 BLASTN 662 1e−46 85 2070 16 700745009H1 SOYMON013 g609224 BLASTN 662 1e−46 85 2071 16 701206618H1 SOYMON035 g16508 BLASTN 663 1e−46 91 2072 16 701131927H1 SOYMON038 g497899 BLASTN 666 1e−46 87 2073 16 701119069H1 SOYMON037 g497899 BLASTN 666 1e−46 87 2074 16 701106908H1 SOYMON036 g497899 BLASTN 666 1e−46 87 2075 16 701103063H1 SOYMON028 g497899 BLASTN 666 1e−46 87 2076 16 700686659H1 SOYMON008 g166873 BLASTN 667 1e−46 86 2077 16 701040620H1 SOYMON029 g16508 BLASTN 669 1e−46 86 2078 16 701012134H1 SOYMON019 g497899 BLASTN 669 1e−46 85 2079 16 700957508H1 SOYMON022 g1724103 BLASTN 669 1e−46 82 2080 16 701008548H1 SOYMON019 g497899 BLASTN 500 1e−45 87 2081 16 700978303H1 SOYMON009 g497899 BLASTN 506 1e−45 87 2082 16 701119990H1 SOYMON037 g497899 BLASTN 512 1e−45 87 2083 16 700646222H1 SOYMON012 g16508 BLASTN 526 1e−45 85 2084 16 701040631H1 SOYMON029 g609556 BLASTN 542 1e−45 87 2085 16 700894009H1 SOYMON024 g497899 BLASTN 646 1e−45 87 2086 16 700908702H1 SOYMON022 g609224 BLASTN 646 1e−45 85 2087 16 700838642H1 SOYMON020 g609224 BLASTN 646 1e−45 85 2088 16 700832089H1 SOYMON019 g609224 BLASTN 646 1e−45 85 2089 16 700978279H1 SOYMON009 g609224 BLASTN 647 1e−45 80 2090 16 700555058H1 SOYMON001 g16508 BLASTN 649 1e−45 84 2091 16 700906478H1 SOYMON022 g497899 BLASTN 651 1e−45 87 2092 16 700897237H1 SOYMON027 g16508 BLASTN 653 1e−45 86 2093 16 701125786H1 SOYMON037 g16508 BLASTN 654 1e−45 84 2094 16 700562836H1 SOYMON002 g16508 BLASTN 655 1e−45 84 2095 16 700665969H1 SOYMON005 g497899 BLASTN 656 1e−45 87 2096 16 701009838H1 SOYMON019 g497899 BLASTN 656 1e−45 87 2097 16 701211770H1 SOYMON035 g16508 BLASTN 657 1e−45 84 2098 16 700654550H1 SOYMON004 g16508 BLASTN 657 1e−45 83 2099 16 700567949H1 SOYMON002 g16508 BLASTN 657 1e−45 84 2100 16 701048175H1 SOYMON032 g16508 BLASTN 658 1e−45 91 2101 16 700901037H1 SOYMON027 g16508 BLASTN 658 1e−45 86 2102 16 700656917H1 SOYMON004 g1724103 BLASTN 407 1e−44 81 2103 16 700565684H1 SOYMON002 g16508 BLASTN 463 1e−44 86 2104 16 701002808H1 SOYMON019 g497899 BLASTN 503 1e−44 87 2105 16 701107115H1 SOYMON036 g497899 BLASTN 507 1e−44 87 2106 16 701122669H1 SOYMON037 g16508 BLASTN 636 1e−44 82 2107 16 700894409H1 SOYMON024 g497899 BLASTN 636 1e−44 87 2108 16 700848374H1 SOYMON021 g497899 BLASTN 636 1e−44 87 2109 16 700902420H1 SOYMON027 g497899 BLASTN 636 1e−44 87 2110 16 700747450H1 SOYMON013 g16508 BLASTN 638 1e−44 91 2111 16 701125869H1 SOYMON037 g16508 BLASTN 638 1e−44 91 2112 16 700731302H1 SOYMON010 g16508 BLASTN 639 1e−44 85 2113 16 701099068H1 SOYMON028 g2305013 BLASTN 640 1e−44 81 2114 16 700975556H1 SOYMON009 g450548 BLASTN 641 1e−44 84 2115 16 700889660H1 SOYMON024 g16508 BLASTN 642 1e−44 86 2116 16 700951744H1 SOYMON022 g16508 BLASTN 642 1e−44 86 2117 16 700750251H1 SOYMON013 g609224 BLASTN 643 1e−44 86 2118 16 700834611H1 SOYMON019 g16508 BLASTN 643 1e−44 91 2119 16 700565356H1 SOYMON002 g16508 BLASTN 643 1e−44 80 2120 16 700673803H1 SOYMON007 g16508 BLASTN 644 1e−44 84 2121 16 700958721H1 SOYMON022 g16508 BLASTN 645 1e−44 86 2122 16 700985717H1 SOYMON009 g16508 BLASTN 646 1e−44 82 2123 16 LIB3049-032-Q1-E1-A5 LIB3049 g16508 BLASTN 662 1e−44 76 2124 16 700733454H1 SOYMON010 g609224 BLASTN 432 1e−43 85 2125 16 700987656H1 SOYMON009 g497899 BLASTN 476 1e−43 87 2126 16 700755957H1 SOYMON014 g497899 BLASTN 483 1e−43 86 2127 16 700749331H1 SOYMON013 g2305013 BLASTN 521 1e−43 84 2128 16 700830984H1 SOYMON019 g497899 BLASTN 622 1e−43 86 2129 16 700906176H1 SOYMON022 g16508 BLASTN 623 1e−43 86 2130 16 701100070H2 SOYMON028 g16508 BLASTN 623 1e−43 91 2131 16 700954053H1 SOYMON022 g497899 BLASTN 624 1e−43 88 2132 16 700833949H1 SOYMON019 g497899 BLASTN 624 1e−43 88 2133 16 700976710H1 SOYMON009 g2305013 BLASTN 625 1e−43 79 2134 16 701117925H2 SOYMON037 g16508 BLASTN 625 1e−43 84 2135 16 700748825H1 SOYMON013 g16508 BLASTN 626 1e−43 84 2136 16 700899651H1 SOYMON027 g16508 BLASTN 626 1e−43 84 2137 16 700746739H1 SOYMON013 g497899 BLASTN 626 1e−43 88 2138 16 701009051H1 SOYMON019 g16508 BLASTN 628 1e−43 91 2139 16 700754423H1 SOYMON014 g16508 BLASTN 628 1e−43 91 2140 16 700760701H1 SOYMON015 g16508 BLASTN 628 1e−43 83 2141 16 701098124H1 SOYMON028 g2305013 BLASTN 628 1e−43 78 2142 16 700984979H1 SOYMON009 g16508 BLASTN 628 1e−43 91 2143 16 700565262H1 SOYMON002 g497899 BLASTN 629 1e−43 86 2144 16 700954979H1 SOYMON022 g497899 BLASTN 630 1e−43 87 2145 16 700834601H1 SOYMON019 g497899 BLASTN 630 1e−43 87 2146 16 700953156H1 SOYMON022 g609224 BLASTN 630 1e−43 85 2147 16 700865217H1 SOYMON016 g16508 BLASTN 631 1e−43 77 2148 16 700852947H1 SOYMON023 g16508 BLASTN 631 1e−43 84 2149 16 700943547H1 SOYMON024 g497899 BLASTN 631 1e−43 86 2150 16 701139277H1 SOYMON038 g609224 BLASTN 632 1e−43 82 2151 16 700900155H1 SOYMON027 g429105 BLASTN 264 1e−42 81 2152 16 700970581H1 SOYMON005 g497899 BLASTN 296 1e−42 86 2153 16 700654232H1 SOYMON003 g497899 BLASTN 348 1e−42 88 2154 16 700743608H1 SOYMON012 g497899 BLASTN 354 1e−42 86 2155 16 700986620H1 SOYMON009 g609224 BLASTN 360 1e−42 84 2156 16 700562590H1 SOYMON002 g16508 BLASTN 476 1e−42 83 2157 16 701119690H1 SOYMON037 g167961 BLASTN 506 1e−42 84 2158 16 700951730H1 SOYMON022 g497899 BLASTN 507 1e−42 87 2159 16 701004525H1 SOYMON019 g16508 BLASTN 516 1e−42 91 2160 16 700892637H1 SOYMON024 g16508 BLASTN 523 1e−42 81 2161 16 700560679H1 SOYMON001 g16508 BLASTN 526 1e−42 83 2162 16 700897888H1 SOYMON027 g16508 BLASTN 611 1e−42 86 2163 16 700547973H1 SOYMON001 g497899 BLASTN 611 1e−42 84 2164 16 701012166H1 SOYMON019 g497899 BLASTN 612 1e−42 87 2165 16 700747550H1 SOYMON013 g16508 BLASTN 613 1e−42 84 2166 16 700946136H1 SOYMON024 g16508 BLASTN 615 1e−42 85 2167 16 700665932H1 SOYMON005 g16508 BLASTN 616 1e−42 84 2168 16 701010969H1 SOYMON019 g16508 BLASTN 618 1e−42 84 2169 16 700962515H1 SOYMON022 g16508 BLASTN 618 1e−42 91 2170 16 700895685H1 SOYMON027 g429107 BLASTN 618 1e−42 77 2171 16 700752015H1 SOYMON014 g16508 BLASTN 620 1e−42 84 2172 16 701015704H1 SOYMON038 g16508 BLASTN 620 1e−42 85 2173 16 700966710H1 SOYMON028 g16508 BLASTN 620 1e−42 83 2174 16 701004268H1 SOYMON019 g16508 BLASTN 620 1e−42 86 2175 16 701138901H1 SOYMON038 g497899 BLASTN 285 1e−41 85 2176 16 700648891H1 SOYMON003 g16508 BLASTN 481 1e−41 84 2177 16 700741421H1 SOYMON012 g609224 BLASTN 511 1e−41 82 2178 16 700555653H1 SOYMON001 g166873 BLASTN 512 1e−41 85 2179 16 700958314H1 SOYMON022 g497899 BLASTN 599 1e−41 87 2180 16 700960055H1 SOYMON022 g497899 BLASTN 599 1e−41 87 2181 16 700901925H1 SOYMON027 g16508 BLASTN 599 1e−41 84 2182 16 700741987H1 SOYMON012 g497899 BLASTN 599 1e−41 87 2183 16 700662837H1 SOYMON005 g497899 BLASTN 599 1e−41 87 2184 16 701060927H1 SOYMON033 g497899 BLASTN 599 1e−41 87 2185 16 700562973H1 SOYMON002 g16508 BLASTN 601 1e−41 87 2186 16 700754586H1 SOYMON014 g16508 BLASTN 603 1e−41 84 2187 16 701132674H1 SOYMON038 g16508 BLASTN 604 1e−41 85 2188 16 701010012H2 SOYMON019 g609224 BLASTN 604 1e−41 84 2189 16 701123643H1 SOYMON037 g16508 BLASTN 604 1e−41 85 2190 16 700894196H1 SOYMON024 g16508 BLASTN 604 1e−41 85 2191 16 700899248H1 SOYMON027 g16508 BLASTN 604 1e−41 85 2192 16 701207680H1 SOYMON035 g16508 BLASTN 605 1e−41 84 2193 16 700899210H1 SOYMON027 g16508 BLASTN 605 1e−41 84 2194 16 700898762H1 SOYMON027 g16508 BLASTN 605 1e−41 87 2195 16 700983636H1 SOYMON009 g497899 BLASTN 606 1e−41 87 2196 16 700845511H1 SOYMON021 g16508 BLASTN 606 1e−41 89 2197 16 700563416H1 SOYMON002 g16508 BLASTN 606 1e−41 83 2198 16 701004041H1 SOYMON019 g16508 BLASTN 607 1e−41 84 2199 16 701097184H1 SOYMON028 g16508 BLASTN 607 1e−41 87 2200 16 700750691H1 SOYMON014 g609224 BLASTN 608 1e−41 84 2201 16 701208091H1 SOYMON035 g16508 BLASTN 608 1e−41 84 2202 16 700973888H1 SOYMON005 g609224 BLASTN 609 1e−41 85 2203 16 700831972H1 SOYMON019 g16508 BLASTN 609 1e−41 85 2204 16 701010090H2 SOYMON019 g16508 BLASTN 610 1e−41 87 2205 16 700890067H1 SOYMON024 g16508 BLASTN 610 1e−41 87 2206 16 700833227H1 SOYMON019 g16508 BLASTN 610 1e−41 85 2207 16 700563171H1 SOYMON002 g16508 BLASTN 370 1e−40 90 2208 16 701121457H1 SOYMON037 g609224 BLASTN 372 1e−40 85 2209 16 700566851H1 SOYMON002 g16508 BLASTN 402 1e−40 82 2210 16 700753221H1 SOYMON014 g609224 BLASTN 422 1e−40 84 2211 16 700749039H1 SOYMON013 g16508 BLASTN 475 1e−40 84 2212 16 700558940H1 SOYMON001 g16508 BLASTN 493 1e−40 80 2213 16 700757695H1 SOYMON015 g16508 BLASTN 526 1e−40 83 2214 16 701102262H1 SOYMON028 g2305013 BLASTN 587 1e−40 81 2215 16 700747484H1 SOYMON013 g16508 BLASTN 587 1e−40 85 2216 16 700891875H1 SOYMON024 g16508 BLASTN 587 1e−40 85 2217 16 700894439H1 SOYMON024 g497899 BLASTN 588 1e−40 88 2218 16 701062577H1 SOYMON033 g16508 BLASTN 588 1e−40 87 2219 16 700964366H1 SOYMON022 g16508 BLASTN 589 1e−40 85 2220 16 701210016H1 SOYMON035 g16508 BLASTN 589 1e−40 83 2221 16 700901573H1 SOYMON027 g16508 BLASTN 589 1e−40 85 2222 16 700750533H1 SOYMON014 g16508 BLASTN 589 1e−40 85 2223 16 700888820H1 SOYMON024 g497899 BLASTN 589 1e−40 87 2224 16 700891965H1 SOYMON024 g497899 BLASTN 589 1e−40 87 2225 16 700865001H1 SOYMON016 g16508 BLASTN 589 1e−40 85 2226 16 700654923H1 SOYMON004 g16508 BLASTN 590 1e−40 84 2227 16 700746169H1 SOYMON013 g16508 BLASTN 590 1e−40 84 2228 16 700745792H1 SOYMON013 g16508 BLASTN 591 1e−40 85 2229 16 700966953H1 SOYMON029 g16508 BLASTN 591 1e−40 85 2230 16 701141657H1 SOYMON038 g16508 BLASTN 592 1e−40 86 2231 16 700943078H1 SOYMON024 g16508 BLASTN 598 1e−40 85 2232 16 701137073H1 SOYMON038 g16508 BLASTN 598 1e−40 90 2233 16 700966730H1 SOYMON028 g16508 BLASTN 598 1e−40 84 2234 16 701137731H1 SOYMON038 g16508 BLASTN 371 1e−39 84 2235 16 701044050H1 SOYMON032 g497899 BLASTN 428 1e−39 89 2236 16 700993309H1 SOYMON011 g497899 BLASTN 434 1e−39 88 2237 16 700874483H1 SOYMON018 g497899 BLASTN 434 1e−39 87 2238 16 701014866H1 SOYMON019 g16508 BLASTN 454 1e−39 89 2239 16 700894603H1 SOYMON024 g16508 BLASTN 454 1e−39 85 2240 16 700958677H1 SOYMON022 g497899 BLASTN 455 1e−39 87 2241 16 700754001H1 SOYMON014 g609224 BLASTN 518 1e−39 85 2242 16 700946430H1 SOYMON024 g497899 BLASTN 578 1e−39 87 2243 16 700729892H1 SOYMON009 g497899 BLASTN 578 1e−39 87 2244 16 700753230H1 SOYMON014 g497899 BLASTN 578 1e−39 87 2245 16 700562584H1 SOYMON002 g16508 BLASTN 578 1e−39 82 2246 16 700842880H1 SOYMON020 g16508 BLASTN 580 1e−39 82 2247 16 700852561H1 SOYMON023 g16508 BLASTN 582 1e−39 85 2248 16 700941290H1 SOYMON024 g497899 BLASTN 583 1e−39 87 2249 16 700750361H1 SOYMON013 g497899 BLASTN 584 1e−39 80 2250 16 700841915H1 SOYMON020 g16508 BLASTN 585 1e−39 85 2251 16 700944462H1 SOYMON024 g16508 BLASTN 585 1e−39 85 2252 16 700548174H1 SOYMON002 g16508 BLASTN 342 1e−38 82 2253 16 700942324H1 SOYMON024 g497899 BLASTN 403 1e−38 87 2254 16 700788450H1 SOYMON011 g497899 BLASTN 416 1e−38 86 2255 16 701127954H1 SOYMON037 g16508 BLASTN 449 1e−38 76 2256 16 700742223H1 SOYMON012 g1724103 BLASTN 451 1e−38 77 2257 16 701102722H1 SOYMON028 g16508 BLASTN 466 1e−38 92 2258 16 701046957H1 SOYMON032 g16508 BLASTN 517 1e−38 85 2259 16 700836169H1 SOYMON019 g497899 BLASTN 517 1e−38 87 2260 16 700697967H1 SOYMON015 g16508 BLASTN 521 1e−38 84 2261 16 700678867H1 SOYMON007 g609224 BLASTN 566 1e−38 82 2262 16 700838753H1 SOYMON020 g16508 BLASTN 567 1e−38 85 2263 16 700726468H1 SOYMON009 g16508 BLASTN 570 1e−38 85 2264 16 701014631H1 SOYMON019 g497899 BLASTN 571 1e−38 87 2265 16 700728719H1 SOYMON009 g497899 BLASTN 573 1e−38 87 2266 16 700962582H1 SOYMON022 g497899 BLASTN 573 1e−38 87 2267 16 701123183H1 SOYMON037 g16508 BLASTN 573 1e−38 84 2268 16 700752347H1 SOYMON014 g16508 BLASTN 574 1e−38 87 2269 16 701103315H1 SOYMON028 g16508 BLASTN 394 1e−37 86 2270 16 700750955H1 SOYMON014 g497899 BLASTN 414 1e−37 88 2271 16 700867493H1 SOYMON016 g609224 BLASTN 433 1e−37 85 2272 16 701054760H1 SOYMON032 g16508 BLASTN 475 1e−37 85 2273 16 700900903H1 SOYMON027 g16508 BLASTN 501 1e−37 84 2274 16 701102294H1 SOYMON028 g16508 BLASTN 502 1e−37 87 2275 16 701124034H1 SOYMON037 g16508 BLASTN 512 1e−37 82 2276 16 700981476H1 SOYMON009 g16508 BLASTN 526 1e−37 83 2277 16 700903712H1 SOYMON022 g609224 BLASTN 552 1e−37 85 2278 16 700648751H1 SOYMON003 g1655577 BLASTN 554 1e−37 86 2279 16 700829930H1 SOYMON019 g609224 BLASTN 556 1e−37 84 2280 16 700758078H1 SOYMON015 g16508 BLASTN 556 1e−37 84 2281 16 700754248H1 SOYMON014 g16508 BLASTN 557 1e−37 81 2282 16 700834650H1 SOYMON019 g609224 BLASTN 557 1e−37 85 2283 16 700746279H1 SOYMON013 g16508 BLASTN 558 1e−37 82 2284 16 700869124H1 SOYMON016 g16508 BLASTN 559 1e−37 83 2285 16 701100219H1 SOYMON028 g2305013 BLASTN 560 1e−37 80 2286 16 700992589H1 SOYMON011 g497899 BLASTN 560 1e−37 80 2287 16 700865642H1 SOYMON016 g16508 BLASTN 562 1e−37 84 2288 16 701068835H1 SOYMON034 g429105 BLASTN 321 1e−36 86 2289 16 700746908H1 SOYMON013 g16508 BLASTN 373 1e−36 81 2290 16 700954719H1 SOYMON022 g16508 BLASTN 475 1e−36 86 2291 16 701042790H1 SOYMON029 g497899 BLASTN 489 1e−36 88 2292 16 700835259H1 SOYMON019 g497899 BLASTN 507 1e−36 88 2293 16 700565816H1 SOYMON002 g16508 BLASTN 519 1e−36 82 2294 16 700895811H1 SOYMON027 g609224 BLASTN 540 1e−36 83 2295 16 700975709H1 SOYMON009 g609224 BLASTN 541 1e−36 84 2296 16 700751645H1 SOYMON014 g16508 BLASTN 542 1e−36 85 2297 16 700567087H1 SOYMON002 g16508 BLASTN 542 1e−36 85 2298 16 700893027H1 SOYMON024 g609224 BLASTN 542 1e−36 85 2299 16 700891185H1 SOYMON024 g609224 BLASTN 543 1e−36 84 2300 16 700851376H1 SOYMON023 g16508 BLASTN 543 1e−36 83 2301 16 700898322H1 SOYMON027 g609224 BLASTN 547 1e−36 85 2302 16 700762881H1 SOYMON015 g16508 BLASTN 323 1e−35 85 2303 16 700960023H1 SOYMON022 g16508 BLASTN 370 1e−35 90 2304 16 701136486H1 SOYMON038 g497899 BLASTN 378 1e−35 85 2305 16 700987688H1 SOYMON009 g497899 BLASTN 387 1e−35 83 2306 16 700836508H1 SOYMON020 g16508 BLASTN 431 1e−35 85 2307 16 700990779H1 SOYMON011 g167961 BLASTN 471 1e−35 86 2308 16 700753019H1 SOYMON014 g497899 BLASTN 476 1e−35 83 2309 16 701138490H1 SOYMON038 g16508 BLASTN 481 1e−35 83 2310 16 700751771H1 SOYMON014 g609224 BLASTN 528 1e−35 84 2311 16 701048324H1 SOYMON032 g609224 BLASTN 529 1e−35 84 2312 16 700835739H1 SOYMON019 g16508 BLASTN 532 1e−35 85 2313 16 700986295H1 SOYMON009 g497899 BLASTN 533 1e−35 86 2314 16 701205693H1 SOYMON035 g16508 BLASTN 535 1e−35 84 2315 16 700865674H1 SOYMON016 g609224 BLASTN 536 1e−35 85 2316 16 700943777H1 SOYMON024 g16508 BLASTN 536 1e−35 84 2317 16 700968207H1 SOYMON035 g609224 BLASTN 536 1e−35 85 2318 16 700898370H1 SOYMON027 g609224 BLASTN 536 1e−35 85 2319 16 700987890H1 SOYMON009 g609224 BLASTN 536 1e−35 85 2320 16 700896016H1 SOYMON027 g609224 BLASTN 536 1e−35 85 2321 16 700554440H1 SOYMON001 g16508 BLASTN 283 1e−34 82 2322 16 700789855H2 SOYMON011 g16508 BLASTN 293 1e−34 91 2323 16 701101393H1 SOYMON028 g16508 BLASTN 311 1e−34 84 2324 16 700946335H1 SOYMON024 g16508 BLASTN 323 1e−34 86 2325 16 701207420H1 SOYMON035 g169664 BLASTN 350 1e−34 88 2326 16 700733034H1 SOYMON010 g16508 BLASTN 384 1e−34 81 2327 16 700790653H2 SOYMON011 g497899 BLASTN 399 1e−34 81 2328 16 700901808H1 SOYMON027 g497899 BLASTN 430 1e−34 81 2329 16 700760954H1 SOYMON015 g609224 BLASTN 514 1e−34 84 2330 16 700896206H1 SOYMON027 g16508 BLASTN 516 1e−34 85 2331 16 701039157H1 SOYMON029 g609224 BLASTN 520 1e−34 76 2332 16 700991056H1 SOYMON011 g166873 BLASTN 520 1e−34 80 2333 16 701097173H1 SOYMON028 g609224 BLASTN 521 1e−34 85 2334 16 700900989H1 SOYMON027 g497899 BLASTN 521 1e−34 86 2335 16 700895584H1 SOYMON027 g609224 BLASTN 521 1e−34 85 2336 16 700726668H1 SOYMON009 g16508 BLASTN 521 1e−34 85 2337 16 701011191H1 SOYMON019 g16508 BLASTN 521 1e−34 85 2338 16 701100827H1 SOYMON028 g16508 BLASTN 522 1e−34 84 2339 16 701156732H1 SOYMON031 g16508 BLASTN 522 1e−34 84 2340 16 700889752H1 SOYMON024 g609224 BLASTN 523 1e−34 84 2341 16 701105873H1 SOYMON036 g16508 BLASTN 524 1e−34 87 2342 16 700958346H1 SOYMON022 g497899 BLASTN 525 1e−34 84 2343 16 700962972H1 SOYMON022 g609224 BLASTN 525 1e−34 84 2344 16 701055918H1 SOYMON032 g16508 BLASTN 526 1e−34 85 2345 16 701123061H1 SOYMON037 g16508 BLASTN 526 1e−34 85 2346 16 701109796H1 SOYMON036 g166873 BLASTN 335 1e−33 86 2347 16 701207350H1 SOYMON035 g497899 BLASTN 349 1e−33 89 2348 16 700835692H1 SOYMON019 g16508 BLASTN 377 1e−33 81 2349 16 700729083H1 SOYMON009 g16508 BLASTN 448 1e−33 85 2350 16 LIB3040-049-Q1-E1-E8 LIB3040 g16508 BLASTN 464 1e−33 80 2351 16 700896567H1 SOYMON027 g497899 BLASTN 502 1e−33 81 2352 16 700668032H1 SOYMON006 g609224 BLASTN 504 1e−33 85 2353 16 700895062H1 SOYMON024 g609224 BLASTN 505 1e−33 83 2354 16 700961178H1 SOYMON022 g16508 BLASTN 506 1e−33 84 2355 16 701202590H1 SOYMON035 g497899 BLASTN 508 1e−33 86 2356 16 700754960H1 SOYMON014 g16508 BLASTN 512 1e−33 83 2357 16 700748584H1 SOYMON013 g16508 BLASTN 512 1e−33 84 2358 16 700791669H1 SOYMON011 g609224 BLASTN 512 1e−33 84 2359 16 701210405H1 SOYMON035 g16508 BLASTN 526 1e−33 83 2360 16 LIB3040-054-Q1-E1-D8 LIB3040 g16508 BLASTN 531 1e−33 80 2361 16 700791176H1 SOYMON011 g497899 BLASTN 311 1e−32 87 2362 16 LIB3049-029-Q1-E1-C5 LIB3049 g167961 BLASTN 368 1e−32 73 2363 16 700893458H1 SOYMON024 g497899 BLASTN 380 1e−32 87 2364 16 700829728H1 SOYMON019 g497899 BLASTN 490 1e−32 88 2365 16 700833137H1 SOYMON019 g609224 BLASTN 490 1e−32 85 2366 16 701204592H2 SOYMON035 g609224 BLASTN 493 1e−32 86 2367 16 700834821H1 SOYMON019 g609224 BLASTN 494 1e−32 78 2368 16 700757167H1 SOYMON015 g16508 BLASTN 494 1e−32 85 2369 16 701203730H2 SOYMON035 g16508 BLASTN 496 1e−32 82 2370 16 701044359H1 SOYMON032 g609224 BLASTN 497 1e−32 84 2371 16 701010095H2 SOYMON019 g16508 BLASTN 498 1e−32 86 2372 16 701100059H2 SOYMON028 g16508 BLASTN 499 1e−32 80 2373 16 700844256H1 SOYMON021 g16508 BLASTN 500 1e−32 80 2374 16 700968296H1 SOYMON035 g497899 BLASTN 501 1e−32 85 2375 16 701110404H1 SOYMON036 g16508 BLASTN 333 1e−31 84 2376 16 700869025H1 SOYMON016 g497899 BLASTN 340 1e−31 86 2377 16 700556245H1 SOYMON001 g609224 BLASTN 355 1e−31 84 2378 16 700566166H1 SOYMON002 g726031 BLASTN 402 1e−31 77 2379 16 700941481H1 SOYMON024 g497899 BLASTN 423 1e−31 79 2380 16 700896940H1 SOYMON027 g169664 BLASTN 477 1e−31 85 2381 16 701145429H1 SOYMON031 g609224 BLASTN 478 1e−31 85 2382 16 700946496H1 SOYMON024 g609224 BLASTN 488 1e−31 86 2383 16 700902149H1 SOYMON027 g1655577 BLASTN 488 1e−31 87 2384 16 700653072H1 SOYMON003 g497899 BLASTN 489 1e−31 88 2385 16 701000261H1 SOYMON018 g16508 BLASTN 495 1e−31 78 2386 16 701039358H1 SOYMON029 g609224 BLASTN 320 1e−30 85 2387 16 701156951H1 SOYMON031 g609224 BLASTN 467 1e−30 86 2388 16 701040880H1 SOYMON029 g497899 BLASTN 477 1e−30 89 2389 16 700979632H2 SQYMON009 g609224 BLASTN 490 1e−30 83 2390 16 700983678H1 SOYMON009 g16508 BLASTN 192 1e−29 86 2391 16 700893644H1 SOYMON024 g609224 BLASTN 320 1e−29 85 2392 16 700962437H1 SOYMON022 g609224 BLASTN 364 1e−29 83 2393 16 701150545H1 SOYMON031 g16508 BLASTN 457 1e−29 86 2394 16 700555403H1 SOYMON001 g16508 BLASTN 458 1e−29 90 2395 16 700794307H1 SOYMON017 g16508 BLASTN 460 1e−29 87 2396 16 701137971H1 SOYMON038 g16508 BLASTN 460 1e−29 84 2397 16 700893774H1 SOYMON024 g609224 BLASTN 461 1e−29 86 2398 16 700565272H1 SOYMON002 g609224 BLASTN 464 1e−29 74 2399 16 700667161H1 SOYMON006 g16508 BLASTN 468 1e−29 75 2400 16 700752952H1 SOYMON014 g609224 BLASTN 279 1e−28 78 2401 16 700901534H1 SOYMON027 g2305013 BLASTN 288 1e−28 85 2402 16 701006617H1 SOYMON019 g497899 BLASTN 358 1e−28 79 2403 16 700753459H1 SOYMON014 g609224 BLASTN 446 1e−28 81 2404 16 701157089H1 SOYMON031 g609224 BLASTN 448 1e−28 84 2405 16 700831271H1 SOYMON019 g16508 BLASTN 453 1e−28 87 2406 16 701207325H1 SOYMON035 g167961 BLASTN 251 1e−27 81 2407 16 700566121H1 SOYMON002 g16508 BLASTN 259 1e−27 75 2408 16 701137878H1 SOYMON038 g609224 BLASTN 280 1e−27 85 2409 16 700763957H1 SOYMON019 g497899 BLASTN 430 1e−27 88 2410 16 701015858H1 SOYMON038 g609224 BLASTN 431 1e−27 86 2411 16 700742849H1 SOYMON012 g497899 BLASTN 434 1e−27 88 2412 16 701102648H1 SOYMON028 g16508 BLASTN 436 1e−27 89 2413 16 700940955H1 SOYMON024 g609556 BLASTN 438 1e−27 85 2414 16 700889805H1 SOYMON024 g16508 BLASTN 441 1e−27 89 2415 16 701061930H1 SOYMON033 g16508 BLASTN 315 1e−26 77 2416 16 700901375H1 SOYMON027 g609224 BLASTN 323 1e−26 84 2417 16 700989056H1 SOYMON011 g609224 BLASTN 345 1e−26 82 2418 16 700665988H1 SOYMON005 g960356 BLASTN 420 1e−26 79 2419 16 700893352H1 SOYMON024 g497899 BLASTN 428 1e−26 87 2420 16 701122945H1 SOYMON037 g16508 BLASTN 269 1e−25 76 2421 16 700992929H1 SOYMON011 g16508 BLASTN 328 1e−25 78 2422 16 700557564H1 SOYMON001 g609224 BLASTN 413 1e−25 85 2423 16 701102620H1 SOYMON028 g609224 BLASTN 414 1e−25 82 2424 16 701062321H1 SOYMON033 g497899 BLASTN 267 1e−24 87 2425 16 701062258H1 SOYMON033 g450548 BLASTN 391 1e−24 83 2426 16 701145703H1 SOYMON031 g609224 BLASTN 411 1e−24 85 2427 16 701144943H1 SOYMON031 g450548 BLASTN 421 1e−24 85 2428 16 701134190H1 SOYMON038 g2305013 BLASTN 268 1e−23 87 2429 16 701132116H1 SOYMON038 g450548 BLASTN 385 1e−23 85 2430 16 701144522H1 SOYMON031 g450548 BLASTN 406 1e−23 85 2431 16 700738002H1 SOYMON012 g1655578 BLASTX 205 1e−21 84 2432 16 700756686H1 SOYMON014 g16508 BLASTN 235 1e−21 76 2433 16 700649058H1 SOYMON003 g16508 BLASTN 360 1e−21 85 2434 16 701152050H1 SOYMON031 g497899 BLASTN 233 1e−20 87 2435 16 701156991H1 SOYMON031 g497900 BLASTX 146 1e−19 98 2436 16 700761949H1 SOYMON015 g497900 BLASTX 173 1e−19 98 2437 16 700988163H1 SOYMON009 g609224 BLASTN 243 1e−19 81 2438 16 700666050H1 SOYMON005 g960357 BLASTX 183 1e−18 89 2439 16 700896896H1 SOYMON027 g497900 BLASTX 95 1e−17 82 2440 16 701100249H1 SOYMON028 g497900 BLASTX 104 1e−17 79 2441 16 701061719H1 SOYMON033 g609224 BLASTN 310 1e−17 67 2442 16 700674836H1 SOYMON007 g16508 BLASTN 331 1e−17 84 2443 16 700908822H1 SOYMON022 g169665 BLASTX 171 1e−16 100 2444 16 700898670H1 SOYMON027 g166872 BLASTX 172 1e−16 93 2445 16 700650967H1 SOYMON003 g16845 BLASTX 147 1e−15 100 2446 16 701132185H1 SOYMON038 g16845 BLASTX 151 1e−15 100 2447 16 700648929H1 SOYMON003 g1033190 BLASTX 156 1e−14 96 2448 16 700960809H1 SOYMON022 g16508 BLASTN 168 1e−14 88 2449 16 701101753H1 SOYMON028 g16845 BLASTX 141 1e−13 88 2450 16 700742010H1 SOYMON012 g609224 BLASTN 273 1e−13 88 2451 16 700735080H1 SOYMON010 g166874 BLASTX 121 1e−12 86 2452 16 700979242H1 SOYMON009 g609224 BLASTN 165 1e−12 81 2453 16 700740838H1 SOYMON012 g609224 BLASTN 253 1e−12 88 2454 16 700832383H1 SOYMON019 g16845 BLASTX 82 1e−11 71 2455 16 700830938H1 SOYMON019 g16845 BLASTX 92 1e−11 90 2456 16 700564686H1 SOYMON002 g1655578 BLASTX 95 1e−11 97 2457 16 700753178H1 SOYMON014 g16845 BLASTX 107 1e−10 78 2458 16 700563834H1 SOYMON002 g497900 BLASTX 124 1e−10 100 2459 16 700897739H1 SOYMON027 g16845 BLASTX 125 1e−10 86 2460 16 700725722H1 SOYMON009 g609225 BLASTX 125 1e−10 92 2461 16 701210357H1 SOYMON035 g497900 BLASTX 128 1e−10 100 2462 16 700833654H1 SOYMON019 g609224 BLASTN 243 1e−10 87 2463 16 701010334H1 SOYMON019 g609224 BLASTN 258 1e−10 86 2464 16 700567622H1 SOYMON002 g166874 BLASTX 93 1e−9 100 2465 16 700647933H1 SOYMON003 g609225 BLASTX 120 1e−9 81 2466 16 700742255H1 SOYMON012 g429107 BLASTN 160 1e−9 86 2467 16 700981506H1 SOYMON009 g609224 BLASTN 248 1e−9 88 2468 16 700962044H1 SOYMON022 g497900 BLASTX 113 1e−8 100 2469 16 701039604H1 SOYMON029 g497900 BLASTX 113 1e−8 100 2470 16 701009941H2 SOYMON019 g609225 BLASTX 114 1e−8 70 2471 16 700976956H1 SOYMON009 g609225 BLASTX 118 1e−8 96 2472 18138 701120722H1 SOYMON037 g169664 BLASTN 514 1e−33 92 2473 18138 700946044H1 SOYMON024 g169664 BLASTN 437 1e−26 90 2474 18138 700664443H1 SOYMON005 g17262 BLASTX 162 1e−16 88 2475 18138 700665162H1 SOYMON005 g17262 BLASTX 162 1e−16 88 2476 18138 701143667H1 SOYMON038 g16961 BLASTX 123 1e−11 95 2477 18138 701099150H1 SOYMON028 g16961 BLASTX 112 1e−9 90 2478 27686 700909141H1 SOYMON022 g726027 BLASTN 723 1e−51 84 2479 27686 701145458H1 SOYMON031 g726027 BLASTN 694 1e−49 86

ADENOSYLMETHIONINE DECARBOXYLASE (EC 4.1.1.50) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 430 -700151703 700151703H1 SATMON007 g1532072 BLASTN 717 1e−50 91 431 -700165608 700165608H1 SATMON013 g1532072 BLASTN 605 1e−41 75 432 -700166868 700166868H1 SATMON013 g1532072 BLASTN 593 1e−40 90 433 -700242148 700242148H1 SATMON010 g1532048 BLASTX 142 1e−12 88 434 -700354923 700354923H1 SATMON024 g1532072 BLASTN 887 1e−74 89 435 -700422279 700422279H1 SATMONN01 g1532072 BLASTN 623 1e−61 88 436 -700455682 700455682H1 SATMON029 g1532047 BLASTN 681 1e−47 77 437 -700477645 700477645H1 SATMON025 g1532048 BLASTX 90 1e−12 71 438 -700550509 700550509H1 SATMON022 g1532047 BLASTN 399 1e−41 80 439 -700572502 700572502H1 SATMON030 g1532048 BLASTX 93 1e−20 70 440 -700618258 700618258H1 SATMON033 g1532072 BLASTN 368 1e−67 92 441 -701165456 701165456H1 SATMONN04 g1532072 BLASTN 436 1e−26 71 442 -L30622289 LIB3062-004-Q1-K1-E10 LIB3062 g1532047 BLASTN 406 1e−26 82 443 -L30623185 LIB3062-026-Q1-K1-E2 LIB3062 g1532072 BLASTN 338 1e−19 62 444 -L30625601 LIB3062-033-Q1-K1-A9 LIB3062 g1403043 BLASTN 758 1e−54 68 445 -L30661842 LIB3066-009-Q1-K1-E4 LIB3066 g1532072 BLASTN 398 1e−22 86 446 -L30681932 LIB3068-020-Q1-K1-C8 LIB3068 g1532072 BLASTN 205 1e−28 89 447 -L30687176 LIB3068-059-Q1-K1-C1 LIB3068 g1532072 BLASTN 216 1e−20 87 448 -L30692075 LIB3069-004-Q1-K1-G11 LIB3069 g1532072 BLASTN 619 1e−40 90 449 -L30783194 LIB3078-052-Q1-K1-E4 LIB3078 g1532072 BLASTN 447 1e−40 76 450 -L30792336 LIB3079-021-Q1-K1-E12 LIB3079 g1532072 BLASTN 217 1e−13 80 451 1471 700106762H1 SATMON010 g1532072 BLASTN 323 1e−59 84 452 1471 701163744H1 SATMONN04 g1532072 BLASTN 220 1e−41 86 453 1471 LIB3059-023-Q1-K1-A11 LIB3059 g1532072 BLASTN 340 1e−39 80 454 1471 700574530H1 SATMON030 g1532072 BLASTN 247 1e−34 82 455 1471 LIB83-003-Q1-E1-G1 LIB83 g1532072 BLASTN 239 1e−24 80 456 1471 701161147H1 SATMONN04 g1532072 BLASTN 239 1e−16 80 457 1471 700477336H1 SATMON025 g1532072 BLASTN 235 1e−10 91 458 16729 700169502H1 SATMON013 g1403043 BLASTN 456 1e−28 65 459 16729 700088201H1 SATMON011 g1532048 BLASTX 123 1e−22 60 460 16729 700165440H1 SATMON013 g1532048 BLASTX 162 1e−15 65 461 16866 700072905H1 SATMON007 g1532047 BLASTN 368 1e−19 71 462 16866 700350558H1 SATMON023 g1532047 BLASTN 301 1e−14 70 463 16866 700088370H1 SATMON011 g1532047 BLASTN 306 1e−14 70 464 2324 LIB3066-042-Q1-K1-H2 LIB3066 g1403043 BLASTN 1344 1e−103 79 465 2324 LIB3059-01 1-Q1-K1-B2 LIB3059 g1403043 BLASTN 1351 1e−103 78 466 2324 LIB3059-042-Q1-K1-B7 LIB3059 g1403043 BLASTN 1230 1e−100 80 467 2324 LIB3062-036-Q1-K1-F2 LIB3062 g1532072 BLASTN 1097 1e−91 80 468 2324 LIB3069-020-Q1-K1-A5 LIB3069 g1403043 BLASTN 925 1e−82 80 469 2324 LIB189-023-Q1-E1-H3 LIB189 g1532072 BLASTN 1085 1e−81 78 470 2324 700266088H1 SATMON017 g1532047 BLASTN 998 1e−74 80 471 2324 LIB3062-026-Q1-K1-E6 LIB3062 g1532047 BLASTN 982 1e−72 77 472 2324 700083335H1 SATMON011 g1403043 BLASTN 960 1e−71 79 473 2324 LIB189-015-Q1-E1-D3 LIB189 g1532047 BLASTN 970 1e−71 77 474 2324 LIB3079-013-Q1-K1-B3 LIB3079 g1403043 BLASTN 953 1e−70 81 475 2324 700262129H1 SATMON017 g1403043 BLASTN 936 1e−69 79 476 2324 700265667H1 SATMON017 g1403043 BLASTN 694 1e−66 81 477 2324 700807255H1 SATMON036 g1532072 BLASTN 850 1e−62 81 478 2324 700196129H1 SATMON014 g1403043 BLASTN 853 1e−62 83 479 2324 LIB3066-042-Q1-K1-H1 LIB3066 g1403043 BLASTN 842 1e−61 80 480 2324 700458321H1 SATMON029 g1403043 BLASTN 501 1e−58 80 481 2324 700197643H1 SATMON014 g1403043 BLASTN 461 1e−57 80 482 2324 LIB143-029-Q1-E1-H3 LIB143 g1532072 BLASTN 786 1e−56 78 483 2324 700197842H1 SATMON014 g1403043 BLASTN 688 1e−48 85 484 2324 700196807H1 SATMON014 g1532072 BLASTN 678 1e−47 75 485 2324 700263712H1 SATMON017 g1532072 BLASTN 403 1e−45 75 486 2324 LIB143-006-Q1-E1-F9 LIB143 g1403043 BLASTN 324 1e−42 83 487 2324 700267496H1 SATMON017 g1532047 BLASTN 594 1e−40 83 488 2324 700172551H1 SATMON013 g1532072 BLASTN 546 1e−36 74 489 2324 700211893H1 SATMON016 g1532047 BLASTN 542 1e−35 82 490 2324 700264065H1 SATMON017 g1532047 BLASTN 515 1e−34 83 491 2324 700465305H1 SATMON025 g1532073 BLASTX 148 1e−26 61 492 2324 700455052H1 SATMON029 g1403043 BLASTN 274 1e−26 79 493 2324 700473305H1 SATMON025 g1403043 BLASTN 446 1e−26 74 494 2324 700475851H1 SATMON025 g1532047 BLASTN 333 1e−18 78 495 2324 700263111H1 SATMON017 g1532073 BLASTX 165 1e−15 72 496 2324 700465705H1 SATMON025 g1403044 BLASTX 78 1e−8 76 497 3185 700264424H1 SATMON017 g1403043 BLASTN 469 1e−41 81 498 3185 LIB143-007-Q1-E1-H2 LIB143 g1403043 BLASTN 464 1e−36 80 499 3185 700262548H1 SATMON017 g1532047 BLASTN 368 1e−34 80 500 3185 700263845H1 SATMON017 g1403043 BLASTN 455 1e−34 81 501 3185 LIB3068-025-Q1-K1-G10 LIB3068 g1403043 BLASTN 288 1e−32 81 502 3185 700264569H1 SATMON017 g1403043 BLASTN 469 1e−32 82 503 3185 700243571H1 SATMON010 g1532047 BLASTN 461 1e−28 77 504 3185 700265453H1 SATMON017 g1532047 BLASTN 269 1e−27 84 505 3185 700267779H1 SATMON017 g1532047 BLASTN 257 1e−26 85 506 3185 700382373H1 SATMON024 g1532047 BLASTN 255 1e−24 86 507 3185 700258727H1 SATMON017 g1532047 BLASTN 255 1e−24 84 508 3185 LIB3069-018-Q1-K1-F5 LIB3069 g1532047 BLASTN 255 1e−21 77 509 3185 LIB3066-038-Q1-K1-C2 LIB3066 g1403043 BLASTN 242 1e−17 68 510 3185 700334654H1 SATMON019 g1532047 BLASTN 248 1e−17 90 511 3185 700262830H1 SATMON017 g1403043 BLASTN 276 1e−12 78 512 3185 700264727H1 SATMON017 g1532047 BLASTN 255 1e−10 92 513 3185 700238407H1 SATMON010 g1532047 BLASTN 255 1e−10 92 514 3185 700441952H1 SATMON026 g1532047 BLASTN 255 1e−10 92 515 3185 700268188H1 SATMON017 g1532047 BLASTN 255 1e−10 92 516 3185 700801627H1 SATMON036 g1532047 BLASTN 255 1e−10 92 517 3185 700261609H1 SATMON017 g1532047 BLASTN 255 1e−10 92 518 3185 700258510H1 SATMON017 g1532047 BLASTN 255 1e−10 92 519 3185 700258779H1 SATMON017 g1532047 BLASTN 255 1e−10 92 520 3185 700256838H1 SATMON017 g1532047 BLASTN 255 1e−10 92 521 3185 700263361H1 SATMON017 g1532047 BLASTN 255 1e−10 92 522 3185 700257102H1 SATMON017 g1532047 BLASTN 255 1e−10 92 523 3185 700239452H1 SATMON010 g1532047 BLASTN 245 1e−9 91 524 3185 700262045H1 SATMON017 g1532047 BLASTN 250 1e−9 90 525 8 LIB3066-027-Q1-K1-C6 LIB3066 g1532072 BLASTN 1414 1e−181 97 526 8 LIB3066-048-Q1-K1-C9 LIB3066 g1532072 BLASTN 1715 1e−173 98 527 8 LIB3066-007-Q1-K1-B4 LIB3066 g1532072 BLASTN 2146 1e−170 97 528 8 LIB3066-019-Q1-K1-B9 LIB3066 g1532072 BLASTN 1884 1e−168 98 529 8 LIB148-038-Q1-E1-A3 LIB148 g1532072 BLASTN 1585 1e−164 99 530 8 LIB3068-002-Q1-K1-H5 LIB3068 g1532072 BLASTN 2051 1e−162 98 531 8 LIB3067-035-Q1-K1-E9 LIB3067 g1532072 BLASTN 1594 1e−161 98 532 8 LIB189-020-Q1-E1-E8 LIB189 g1532072 BLASTN 1501 1e−160 98 533 8 LIB3078-057-Q1-K1-C12 LIB3078 g1532072 BLASTN 2026 1e−160 96 534 8 LIB3066-053-Q1-K1-A8 LIB3066 g1532072 BLASTN 1713 1e−159 93 535 8 LIB189-006-Q1-E1-C6 LIB189 g1532072 BLASTN 1676 1e−157 98 536 8 LIB3078-054-Q1-K1-E4 LIB3078 g1532072 BLASTN 1997 1e−157 95 537 8 LIB3069-028-Q1-K1-A11 LIB3069 g1532072 BLASTN 1980 1e−156 96 538 8 LIB3069-045-Q1-K1-H7 LIB3069 g1532072 BLASTN 1989 1e−156 98 539 8 LIB148-025-Q1-E1-F7 LIB148 g1532072 BLASTN 1954 1e−154 97 540 8 LIB189-014-Q1-E1-F11 LIB189 g1532072 BLASTN 1925 1e−151 94 541 8 LIB3060-014-Q1-K1-C5 LIB3060 g1532072 BLASTN 1780 1e−150 96 542 8 LIB148-057-Q1-E1-C2 LIB148 g1532072 BLASTN 1000 1e−149 98 543 8 LIB148-015-Q1-E1-F2 LIB148 g1532072 BLASTN 1886 1e−148 94 544 8 LIB3066-045-Q1-K1-A2 LIB3066 g1532072 BLASTN 914 1e−146 91 545 8 LIB148-064-Q1-E1-A3 LIB148 g1532072 BLASTN 1660 1e−146 93 546 8 LIB3061-047-Q1-K1-G1 LIB3061 g1532072 BLASTN 1867 1e−146 92 547 8 LIB148-040-Q1-E1-F1 LIB148 g1532072 BLASTN 1706 1e−145 94 548 8 LIB3069-012-Q1-K1-B6 LIB3069 g1532072 BLASTN 1630 1e−144 87 549 8 LIB3068-009-Q1-K1-A8 LIB3068 g1532072 BLASTN 1802 1e−141 97 550 8 LIB148-004-Q1-E1-D2 LIB148 g1532072 BLASTN 818 1e−139 91 551 8 LIB3068-002-Q1-K1-A1 LIB3068 g1532072 BLASTN 944 1e−137 95 552 8 LIB3067-035-Q1-K1-H9 LIB3067 g1532072 BLASTN 1701 1e−137 98 553 8 LIB3068-033-Q1-K1-G12 LIB3068 g1532072 BLASTN 1093 1e−130 89 554 8 700572229H1 SATMON030 g1532072 BLASTN 1135 1e−128 99 555 8 LIB3078-052-Q1-K1-E1 LIB3078 g1532072 BLASTN 1235 1e−127 85 556 8 700572579H1 SATMON030 g1532072 BLASTN 1625 1e−126 98 557 8 LIB3066-025-Q1-K1-F2 LIB3066 g1532072 BLASTN 1625 1e−126 100 558 8 LIB3068-048-Q1-K1-F9 LIB3068 g1532072 BLASTN 1538 1e−125 98 559 8 700098413H1 SATMON009 g1532072 BLASTN 1595 1e−124 100 560 8 700573235H1 SATMON030 g1532072 BLASTN 1513 1e−123 98 561 8 700090946H1 SATMON011 g1532072 BLASTN 1585 1e−123 100 562 8 700092465H1 SATMON008 g1532072 BLASTN 1541 1e−122 98 563 8 700074625H1 SATMON007 g1532072 BLASTN 1570 1e−122 100 564 8 LIB3059-007-Q1-K1-C10 LIB3059 g1532072 BLASTN 1401 1e−119 94 565 8 700072828H1 SATMON007 g1532072 BLASTN 1540 1e−119 100 566 8 700619106H1 SATMON034 g1532072 BLASTN 833 1e−118 97 567 8 700074853H1 SATMON007 g1532072 BLASTN 1515 1e−117 100 568 8 700201293H1 SATMON003 g1532072 BLASTN 1517 1e−117 97 569 8 700075896H1 SATMON007 g1532072 BLASTN 1006 1e−115 99 570 8 LIB3059-052-Q1-K1-A1 LIB3059 g1532072 BLASTN 1241 1e−115 92 571 8 700091576H1 SATMON011 g1532072 BLASTN 943 1e−114 98 572 8 LIB3078-015-Q1-K1-D7 LIB3078 g1532072 BLASTN 1218 1e−114 87 573 8 700074733H1 SATMON007 g1532072 BLASTN 1475 1e−114 100 574 8 700381421H1 SATMON023 g1532072 BLASTN 1475 1e−114 100 575 8 700085594H1 SATMON011 g1532072 BLASTN 1477 1e−114 99 576 8 700095883H1 SATMON008 g1532072 BLASTN 1477 1e−114 99 577 8 700338237H1 SATMON020 g1532072 BLASTN 1478 1e−114 99 578 8 700549813H1 SATMON022 g1532072 BLASTN 1481 1e−114 99 579 8 700097935H1 SATMON009 g1532072 BLASTN 1484 1e−114 98 580 8 700572978H1 SATMON030 g1532072 BLASTN 865 1e−113 96 581 8 700196464H1 SATMON014 g1532072 BLASTN 1044 1e−113 92 582 8 700381412H1 SATMON023 g1532072 BLASTN 1168 1e−113 98 583 8 700027839H1 SATMON003 g1532072 BLASTN 1472 1e−113 99 584 8 700623344H1 SATMON034 g1532072 BLASTN 1085 1e−112 94 585 8 700025858H1 SATMON003 g1532072 BLASTN 1455 1e−112 100 586 8 LIB3059-017-Q1-K1-H5 LIB3059 g1532072 BLASTN 1459 1e−112 97 587 8 700475019H1 SATMON025 g1532072 BLASTN 1390 1e−111 100 588 8 700338357H1 SATMON020 g1532072 BLASTN 1440 1e−111 100 589 8 700256796H1 SATMON017 g1532072 BLASTN 1400 1e−110 100 590 8 700071692H1 SATMON007 g1532072 BLASTN 1430 1e−110 98 591 8 700339073H1 SATMON020 g1532072 BLASTN 1357 1e−109 95 592 8 700106916H1 SATMON010 g1532072 BLASTN 1415 1e−109 93 593 8 700468234H1 SATMON025 g1532072 BLASTN 1420 1e−109 100 594 8 700214482H1 SATMON016 g1532072 BLASTN 1425 1e−109 100 595 8 700466437H1 SATMON025 g1532072 BLASTN 1403 1e−108 98 596 8 700205576H1 SATMON003 g1532072 BLASTN 1404 1e−108 98 597 8 700043455H1 SATMON004 g1532072 BLASTN 1405 1e−108 100 598 8 700348439H1 SATMON023 g1532072 BLASTN 1407 1e−108 99 599 8 700093131H1 SATMON008 g1532072 BLASTN 755 1e−107 100 600 8 700571851H1 SATMON030 g1532072 BLASTN 1302 1e−107 99 601 8 700088142H1 SATMON011 g1532072 BLASTN 1392 1e−107 99 602 8 700085934H1 SATMON011 g1532072 BLASTN 1397 1e−107 98 603 8 700028465H1 SATMON003 g1532072 BLASTN 1258 1e−106 98 604 8 700236818H1 SATMON010 g1532072 BLASTN 1380 1e−106 100 605 8 700583691H1 SATMON031 g1532072 BLASTN 1382 1e−106 99 606 8 700095585H1 SATMON008 g1532072 BLASTN 1383 1e−106 94 607 8 700105140H1 SATMON010 g1532072 BLASTN 1375 1e−105 100 608 8 700338486H1 SATMON020 g1532072 BLASTN 893 1e−104 98 609 8 700214178H1 SATMON016 g1532072 BLASTN 1361 1e−104 99 610 8 700090613H1 SATMON011 g1532072 BLASTN 699 1e−103 99 611 8 700576189H1 SATMON030 g1532072 BLASTN 1254 1e−103 93 612 8 700475734H1 SATMON025 g1532072 BLASTN 1256 1e−103 99 613 8 700028218H1 SATMON003 g1532072 BLASTN 1275 1e−103 100 614 8 700088644H1 SATMON011 g1532072 BLASTN 1351 1e−103 98 615 8 700378768H1 SATMON020 g1532072 BLASTN 1067 1e−102 98 616 8 LIB3067-035-Q1-K1-H10 LIB3067 g1532072 BLASTN 1233 1e−102 95 617 8 700043466H1 SATMON004 g1532072 BLASTN 1331 1e−102 97 618 8 LIB148-057-Q1-E1-C3 LIB148 g1532072 BLASTN 1283 1e−101 92 619 8 700217574H1 SATMON016 g1532072 BLASTN 1320 1e−101 100 620 8 700042332H1 SATMON004 g1532072 BLASTN 1321 1e−101 97 621 8 700440926H1 SATMON026 g1532072 BLASTN 1323 1e−101 99 622 8 700552284H1 SATMON022 g1532072 BLASTN 1329 1e−101 97 623 8 700216613H1 SATMON016 g1532072 BLASTN 689 1e−100 97 624 8 LIB3067-054-Q1-K1-F6 LIB3067 g1532072 BLASTN 1030 1e−100 94 625 8 LIB3060-038-Q1-K1-B9 LIB3060 g1532072 BLASTN 1108 1e−100 92 626 8 700218755H1 SATMON011 g1532072 BLASTN 1116 1e−100 99 627 8 700338042H1 SATMON020 g1532072 BLASTN 1270 1e−100 99 628 8 700268009H1 SATMON017 g1532072 BLASTN 1307 1e−100 92 629 8 700578491H1 SATMON031 g1532072 BLASTN 1309 1e−100 97 630 8 700030037H1 SATMON003 g1532072 BLASTN 1310 1e−100 97 631 8 700221406H1 SATMON011 g1532072 BLASTN 1315 1e−100 100 632 8 700157357H1 SATMON012 g1532072 BLASTN 1315 1e−100 98 633 8 700476926H1 SATMON025 g1532072 BLASTN 680 1e−99 97 634 8 700475068H1 SATMON025 g1532072 BLASTN 792 1e−99 98 635 8 700469741H1 SATMON025 g1532072 BLASTN 831 1e−99 99 636 8 700082377H1 SATMON011 g1532072 BLASTN 1081 1e−99 99 637 8 700217143H1 SATMON016 g1532072 BLASTN 1208 1e−99 98 638 8 700339433H1 SATMON020 g1532072 BLASTN 1295 1e−99 100 639 8 700196627H1 SATMON014 g1532072 BLASTN 1295 1e−99 100 640 8 700259354H1 SATMON017 g1532072 BLASTN 1305 1e−99 92 641 8 700610842H1 SATMON022 g1532072 BLASTN 843 1e−98 97 642 8 700218059H1 SATMON016 g1532072 BLASTN 1036 1e−98 99 643 8 700421727H1 SATMONN01 g1532072 BLASTN 1172 1e−98 97 644 8 700158660H1 SATMON012 g1532072 BLASTN 1285 1e−98 100 645 8 700806856H1 SATMON036 g1532072 BLASTN 1194 1e−97 97 646 8 700583512H1 SATMON031 g1532072 BLASTN 1219 1e−97 97 647 8 700025502H1 SATMON004 g1532072 BLASTN 1276 1e−97 99 648 8 700159562H1 SATMON012 g1532072 BLASTN 1277 1e−97 99 649 8 700223731H1 SATMON011 g1532072 BLASTN 1277 1e−97 99 650 8 700156494H1 SATMON012 g1532072 BLASTN 1280 1e−97 100 651 8 700160533H1 SATMON012 g1532072 BLASTN 1281 1e−97 97 652 8 LIB3066-055-Q1-K1-D8 LIB3066 g1532072 BLASTN 672 1e−96 99 653 8 700405152H1 SATMON028 g1532072 BLASTN 828 1e−96 98 654 8 LIB148-040-Q1-E1-A8 LIB148 g1532072 BLASTN 1268 1e−96 83 655 8 700438437H1 SATMON026 g1532072 BLASTN 1246 1e−95 99 656 8 700267245H1 SATMON017 g1532072 BLASTN 1248 1e−95 93 657 8 700156129H2 SATMON007 g1532072 BLASTN 1250 1e−95 100 658 8 700203246H1 SATMON003 g1532072 BLASTN 1250 1e−95 100 659 8 700168339H1 SATMON013 g1532072 BLASTN 915 1e−94 98 660 8 LIB3067-036-Q1-K1-F9 LIB3067 g1532072 BLASTN 1110 1e−94 95 661 8 700193769H1 SATMON014 g1532072 BLASTN 1240 1e−94 100 662 8 700094014H1 SATMON008 g1532072 BLASTN 1241 1e−94 91 663 8 700020311H1 SATMON001 g1532072 BLASTN 1228 1e−93 99 664 8 700195083H1 SATMON014 g1532072 BLASTN 1229 1e−93 98 665 8 700193901H1 SATMON014 g1532072 BLASTN 1229 1e−93 98 666 8 700194639H1 SATMON014 g1532072 BLASTN 1231 1e−93 99 667 8 700350117H1 SATMON023 g1532072 BLASTN 1143 1e−92 99 668 8 700239867H1 SATMON010 g1532072 BLASTN 1210 1e−92 98 669 8 LIB3069-028-Q1-K1-D1 LIB3069 g1532072 BLASTN 1217 1e−92 94 670 8 700355756H1 SATMON024 g1532072 BLASTN 618 1e−91 96 671 8 700265492H1 SATMON017 g1532072 BLASTN 620 1e−91 94 672 8 700579021H1 SATMON031 g1532072 BLASTN 656 1e−91 96 673 8 LIB3079-015-Q1-K1-B11 LIB3079 g1532072 BLASTN 1008 1e−91 83 674 8 700572888H2 SATMON030 g1532072 BLASTN 1159 1e−91 99 675 8 701183990H1 SATMONN06 g1532072 BLASTN 1198 1e−91 93 676 8 700085757H1 SATMON011 g1532072 BLASTN 1200 1e−91 100 677 8 700162756H1 SATMON013 g1532072 BLASTN 1208 1e−91 99 678 8 700193076H1 SATMON014 g1532072 BLASTN 1208 1e−91 99 679 8 700224378H1 SATMON011 g1532072 BLASTN 696 1e−90 99 680 8 700218773H1 SATMON011 g1532072 BLASTN 1187 1e−90 93 681 8 700159338H1 SATMON012 g1532072 BLASTN 1193 1e−90 97 682 8 700169217H1 SATMON013 g1532072 BLASTN 1196 1e−90 99 683 8 700551548H1 SATMON022 g1532072 BLASTN 839 1e−89 96 684 8 700197059H1 SATMON014 g1532072 BLASTN 1025 1e−89 95 685 8 700469834H1 SATMON025 g1532072 BLASTN 650 1e−88 97 686 8 700569778H1 SATMON030 g1532072 BLASTN 682 1e−88 91 687 8 700194845H1 SATMON014 g1532072 BLASTN 1171 1e−88 99 688 8 700445861H1 SATMON027 g1532072 BLASTN 536 1e−87 99 689 8 700100536H1 SATMON009 g1532072 BLASTN 666 1e−87 94 690 8 701163801H1 SATMONN04 g1532072 BLASTN 805 1e−87 94 691 8 LIB3060-035-Q1-K1-E3 LIB3060 g1532072 BLASTN 922 1e−87 95 692 8 700170872H1 SATMON013 g1532072 BLASTN 943 1e−87 97 693 8 700241270H1 SATMON010 g1532072 BLASTN 1066 1e−86 96 694 8 700457981H1 SATMON029 g1532072 BLASTN 1140 1e−86 92 695 8 700158339H1 SATMON012 g1532072 BLASTN 1142 1e−86 99 696 8 700019442H1 SATMON001 g1532072 BLASTN 1145 1e−86 100 697 8 700149885H1 SATMON007 g1532072 BLASTN 1133 1e−85 99 698 8 700018255H1 SATMON001 g1532072 BLASTN 1135 1e−85 100 699 8 700244026H1 SATMON010 g1532072 BLASTN 1044 1e−84 89 700 8 LIB3060-027-Q1-K1-B2 LIB3060 g1532072 BLASTN 1117 1e−84 99 701 8 700170960H1 SATMON013 g1532072 BLASTN 696 1e−83 99 702 8 700267487H1 SATMON017 g1532072 BLASTN 1061 1e−83 92 703 8 700152754H1 SATMON007 g1532072 BLASTN 1107 1e−83 99 704 8 700378260H1 SATMON019 g1532072 BLASTN 1112 1e−83 92 705 8 700455089H1 SATMON029 g1532072 BLASTN 583 1e−82 97 706 8 700167322H1 SATMON013 g1532072 BLASTN 1080 1e−81 98 707 8 LIB3060-035-Q1-K1-H1 LIB3060 g1532072 BLASTN 782 1e−80 94 708 8 700204067H1 SATMON003 g1532072 BLASTN 1011 1e−80 98 709 8 700049271H1 SATMON003 g1532072 BLASTN 627 1e−79 93 710 8 700442065H1 SATMON026 g1532072 BLASTN 1043 1e−78 91 711 8 700244175H1 SATMON010 g1532072 BLASTN 1044 1e−78 90 712 8 700045296H1 SATMON004 g1532072 BLASTN 1041 1e−77 93 713 8 700578391H1 SATMON031 g1532072 BLASTN 624 1e−76 88 714 8 700346136H1 SATMON021 g1532072 BLASTN 751 1e−76 90 715 8 700457852H1 SATMON029 g1532072 BLASTN 777 1e−76 92 716 8 700570111H1 SATMON030 g1532072 BLASTN 814 1e−75 94 717 8 LIB3066-030-Q1-K1-A12 LIB3066 g1532072 BLASTN 922 1e−75 94 718 8 700149657H1 SATMON007 g1532072 BLASTN 1012 1e−75 92 719 8 LIB3060-043-Q1-K1-B4 LIB3060 g1532072 BLASTN 1013 1e−75 97 720 8 700156752H1 SATMON012 g1532072 BLASTN 1013 1e−75 98 721 8 700454114H1 SATMON029 g1532072 BLASTN 455 1e−74 93 722 8 LIB143-053-Q1-E1-E8 LIB143 g1532072 BLASTN 672 1e−74 98 723 8 700029323H1 SATMON003 g1532072 BLASTN 996 1e−74 99 724 8 700156381H1 SATMON007 g1532072 BLASTN 1002 1e−74 97 725 8 700159603H2 SATMON012 g1532072 BLASTN 1003 1e−74 94 726 8 LIB3060-035-Q1-K1-E5 LIB3060 g1532072 BLASTN 603 1e−72 94 727 8 LIB3069-025-Q1-K1-E12 LIB3069 g1532072 BLASTN 734 1e−72 90 728 8 700166680H1 SATMON013 g1532072 BLASTN 971 1e−72 99 729 8 700171123H1 SATMON013 g1532072 BLASTN 973 1e−72 94 730 8 LIB148-042-Q1-E1-G10 LIB148 g1532072 BLASTN 975 1e−72 100 731 8 700612951H1 SATMON033 g1532072 BLASTN 401 1e−71 96 732 8 700265213H1 SATMON017 g1532072 BLASTN 626 1e−71 92 733 8 LIB189-030-Q1-E1-D11 LIB189 g1532072 BLASTN 740 1e−70 92 734 8 700212877H1 SATMON016 g1532072 BLASTN 918 1e−70 93 735 8 700161545H1 SATMON012 g1532072 BLASTN 621 1e−69 99 736 8 700021259H1 SATMON001 g1532072 BLASTN 940 1e−69 93 737 8 LIB3079-015-Q1-K1-C7 LIB3079 g1532072 BLASTN 508 1e−68 96 738 8 LIB3066-055-Q1-K1-F11 LIB3066 g1532072 BLASTN 832 1e−68 85 739 8 700166771H1 SATMON013 g1532072 BLASTN 922 1e−68 88 740 8 LIB3066-003-Q1-K1-E6 LIB3066 g1532072 BLASTN 927 1e−68 86 741 8 700208131H1 SATMON016 g1532047 BLASTN 747 1e−66 85 742 8 700020688H1 SATMON001 g1532072 BLASTN 905 1e−66 90 743 8 700206405H1 SATMON003 g1532072 BLASTN 905 1e−66 100 744 8 LIB3069-042-Q1-K1-C5 LIB3069 g1532072 BLASTN 891 1e−65 99 745 8 700353835H1 SATMON024 g1532072 BLASTN 896 1e−65 96 746 8 700471533H1 SATMON025 g1532072 BLASTN 496 1e−63 99 747 8 700165425H1 SATMON013 g1532072 BLASTN 850 1e−62 100 748 8 LIB3069-054-Q1-K1-C4 LIB3069 g1532072 BLASTN 613 1e−60 87 749 8 700160896H1 SATMON012 g1532072 BLASTN 746 1e−60 95 750 8 LIB3069-042-Q1-K1-A11 LIB3069 g1532072 BLASTN 840 1e−60 98 751 8 700466625H1 SATMON025 g1532072 BLASTN 635 1e−59 86 752 8 700571770H1 SATMON030 g1532072 BLASTN 820 1e−59 84 753 8 LIB143-024-Q1-E1-D2 LIB143 g1532072 BLASTN 813 1e−58 95 754 8 700453677H1 SATMON028 g1532072 BLASTN 508 1e−57 92 755 8 700193979H1 SATMON014 g1532072 BLASTN 790 1e−57 100 756 8 700467828H1 SATMON025 g1532072 BLASTN 779 1e−56 96 757 8 700150072H1 SATMON007 g1532072 BLASTN 776 1e−55 99 758 8 700802869H1 SATMON036 g1532072 BLASTN 702 1e−54 95 759 8 LIB148-051-Q1-E1-B12 LIB148 g1532072 BLASTN 421 1e−53 92 760 8 700075035H1 SATMON007 g1532072 BLASTN 506 1e−53 93 761 8 700471283H1 SATMON025 g1532072 BLASTN 742 1e−53 91 762 8 700166625H1 SATMON013 g1532072 BLASTN 748 1e−53 99 763 8 700019894H1 SATMON001 g1532072 BLASTN 643 1e−51 89 764 8 700204863H1 SATMON003 g1532072 BLASTN 721 1e−51 99 765 8 700431071H1 SATMONN01 g1532072 BLASTN 366 1e−50 94 766 8 700171582H1 SATMON013 g1532072 BLASTN 492 1e−50 99 767 8 700450579H1 SATMON028 g1532072 BLASTN 338 1e−49 97 768 8 700084149H1 SATMON011 g1532072 BLASTN 672 1e−47 98 769 8 700095070H1 SATMON008 g1532072 BLASTN 662 1e−46 98 770 8 700570827H1 SATMON030 g1532072 BLASTN 344 1e−44 83 771 8 LIB3061-057-Q1-K1-G12 LIB3061 g1532072 BLASTN 378 1e−43 75 772 8 700194918H1 SATMON014 g1532072 BLASTN 626 1e−43 89 773 8 700378894H1 SATMON020 g1532072 BLASTN 495 1e−42 97 774 8 700468029H1 SATMON025 g1532072 BLASTN 616 1e−42 98 775 8 LIB143-012-Q1-E1-H8 LIB143 g1532072 BLASTN 637 1e−42 98 776 8 LIB3060-037-Q1-K1-H4 LIB3060 g1532072 BLASTN 638 1e−42 86 777 8 700433327H1 SATMONN01 g1532072 BLASTN 365 1e−41 86 778 8 700623611H1 SATMON034 g1532072 BLASTN 316 1e−39 95 779 8 700166123H1 SATMON013 g1532072 BLASTN 585 1e−39 100 780 8 700616273H1 SATMON033 g1532072 BLASTN 534 1e−38 98 781 8 700464702H1 SATMON025 g1532072 BLASTN 566 1e−38 99 782 8 700338809H1 SATMON020 g1532072 BLASTN 550 1e−37 100 783 8 700092622H1 SATMON008 g1532072 BLASTN 561 1e−37 99 784 8 LIB3060-043-Q1-K1-A10 LIB3060 g1532072 BLASTN 352 1e−36 93 785 8 700265611H1 SATMON017 g1532072 BLASTN 548 1e−36 95 786 8 700100319H1 SATMON009 g1532072 BLASTN 552 1e−36 98 787 8 700092696H1 SATMON008 g1532072 BLASTN 552 1e−36 98 788 8 700076750H1 SATMON007 g1532072 BLASTN 526 1e−35 99 789 8 700082896H1 SATMON011 g1532072 BLASTN 526 1e−35 99 790 8 700075411H1 SATMON007 g1532072 BLASTN 319 1e−34 91 791 8 701178051H1 SATMONN05 g1532072 BLASTN 342 1e−34 94 792 8 700266358H1 SATMON017 g1532072 BLASTN 520 1e−34 95 793 8 700453981H1 SATMON029 g1532072 BLASTN 505 1e−33 96 794 8 700584238H1 SATMON031 g1532072 BLASTN 509 1e−33 91 795 8 700103871H1 SATMON010 g1532072 BLASTN 491 1e−32 99 796 8 700264460H1 SATMON017 g1532072 BLASTN 495 1e−32 95 797 8 700205122H1 SATMON003 g1532072 BLASTN 501 1e−32 99 798 8 700165768H1 SATMON013 g1532072 BLASTN 420 1e−31 98 799 8 700077179H1 SATMON007 g1532072 BLASTN 471 1e−30 98 800 8 700476654H1 SATMON025 g1532072 BLASTN 476 1e−30 98 801 8 700027374H1 SATMON003 g1532072 BLASTN 476 1e−30 98 802 8 700266994H1 SATMON017 g1532072 BLASTN 476 1e−30 98 803 8 700088193H1 SATMON011 g1532072 BLASTN 488 1e−30 97 804 8 700214511H1 SATMON016 g1532072 BLASTN 298 1e−29 93 805 8 700257186H1 SATMON017 g1532072 BLASTN 465 1e−29 95 806 8 700335814H1 SATMON019 g1532072 BLASTN 469 1e−29 93 807 8 700236166H1 SATMON010 g1532072 BLASTN 450 1e−28 95 808 8 700468243H1 SATMON025 g1532072 BLASTN 456 1e−28 98 809 8 700096327H1 SATMON008 g1532072 BLASTN 451 1e−27 98 810 8 700471139H1 SATMON025 g1532072 BLASTN 451 1e−27 98 811 8 700050420H1 SATMON003 g1532072 BLASTN 365 1e−26 92 812 8 700264846H1 SATMON017 g1532072 BLASTN 430 1e−25 94 813 8 700266803H1 SATMON017 g1532072 BLASTN 293 1e−24 75 814 8 700086134H1 SATMON011 g1532072 BLASTN 422 1e−24 97 815 8 700162001H1 SATMON012 g1532072 BLASTN 279 1e−23 96 816 8 700044825H1 SATMON004 g1532072 BLASTN 411 1e−23 98 817 8 700074679H1 SATMON007 g1532072 BLASTN 411 1e−23 98 818 8 700267694H1 SATMON017 g1532072 BLASTN 320 1e−22 92 819 8 700267990H1 SATMON017 g1532072 BLASTN 361 1e−22 99 820 8 700456715H1 SATMON029 g1532072 BLASTN 293 1e−21 97 821 8 700454806H1 SATMON029 g1532072 BLASTN 356 1e−21 98 822 8 700466812H1 SATMON025 g1532072 BLASTN 380 1e−21 95 823 8 700046476H1 SATMON004 g1532072 BLASTN 381 1e−21 97 824 8 700207160H1 SATMON017 g1532072 BLASTN 393 1e−21 96 825 8 700383037H1 SATMON024 g1532072 BLASTN 280 1e−20 98 826 8 700549660H1 SATMON022 g1532072 BLASTN 376 1e−20 97 827 8 LIB3066-004-Q1-K1-G12 LIB3066 g1532072 BLASTN 379 1e−20 91 828 8 700801422H1 SATMON036 g1532072 BLASTN 321 1e−19 97 829 8 700045319H1 SATMON004 g1532072 BLASTN 336 1e−17 99 830 8 700046047H1 SATMON004 g1532072 BLASTN 336 1e−17 99 831 8 700264328H1 SATMON017 g1532072 BLASTN 339 1e−17 95 832 8 700083485H1 SATMON011 g1532072 BLASTN 341 1e−17 99 833 8 700461268H1 SATMON033 g1532072 BLASTN 342 1e−17 95 834 8 700053271H1 SATMON008 g1532072 BLASTN 311 1e−15 98 835 8 700215275H1 SATMON016 g1532072 BLASTN 321 1e−15 98 836 8 700623133H1 SATMON034 g1532072 BLASTN 275 1e−14 75 837 8 700454949H1 SATMON029 g1532072 BLASTN 280 1e−14 99 838 8 700440763H1 SATMON026 g1532072 BLASTN 307 1e−14 95 839 8 LIB3068-022-Q1-K1-C4 LIB3068 g1532072 BLASTN 150 1e−12 94 840 8 700798849H1 SATMON036 g1532072 BLASTN 281 1e−12 98 841 8 700333057H1 SATMON019 g1532072 BLASTN 184 1e−11 91 842 8 700265786H1 SATMON017 g1532072 BLASTN 200 1e−11 90 843 8 700193030H1 SATMON014 g1532072 BLASTN 158 1e−9 94 844 8 700262964H1 SATMON017 g1532047 BLASTN 171 1e−9 81 845 8 LIB3069-023-Q1-K1-B2 LIB3069 g1403043 BLASTN 195 1e−9 84 846 8 700474096H1 SATMON025 g1532072 BLASTN 199 1e−9 97 847 8 700028326H1 SATMON003 g1532072 BLASTN 151 1e−8 96 848 8011 700440327H1 SATMON026 g1532047 BLASTN 292 1e−14 85 849 851 LIB3059-052-Q1-K1-E6 LIB3059 g1403043 BLASTN 1033 1e−85 75 850 851 700090958H1 SATMON011 g1532072 BLASTN 1044 1e−78 80 851 851 700224605H1 SATMON011 g1532072 BLASTN 850 1e−62 80 852 851 700551562H1 SATMON022 g1403043 BLASTN 374 1e−55 79 853 851 700153939H1 SATMON007 g1403043 BLASTN 772 1e−55 80 854 851 700469231H1 SATMON025 g1532072 BLASTN 560 1e−52 76 855 851 700349854H1 SATMON023 g1403043 BLASTN 669 1e−46 81 856 851 701169324H1 SATMONN05 g1403043 BLASTN 669 1e−46 78 857 851 700088319H1 SATMON011 g1532072 BLASTN 492 1e−30 78 2480 -700998660 700998660H1 SOYMON018 g1531764 BLASTN 249 1e−29 90 2481 -GM927 LIB3028-005-Q1-B1-B4 LIB3028 g1421750 BLASTN 261 1e−10 80 2482 13379 700842514H1 SOYMON020 g1915980 BLASTN 575 1e−41 73 2483 13379 701042786H1 SOYMON029 g1421750 BLASTN 377 1e−20 80 2484 15556 701101584H1 SOYMON028 g1421750 BLASTN 526 1e−35 69 2485 15556 701099749H1 SOYMON028 g1155239 BLASTN 453 1e−27 72 2486 16 LIB3051-018-Q1-E1-A7 LIB3051 g1421750 BLASTN 1146 1e−93 81 2487 16 LIB3056-005-Q1-N1-E10 LIB3056 g1421750 BLASTN 944 1e−88 80 2488 16 LIB3051-101-Q1-K1-C7 LIB3051 g1421750 BLASTN 1135 1e−85 80 2489 16 LIB3056-001-Q1-B1-H12 LIB3056 g1421750 BLASTN 752 1e−82 81 2490 16 700661334H1 SOYMON005 g1421750 BLASTN 1092 1e−82 80 2491 16 LIB3056-012-Q1-N1-B12 LIB3056 g1421750 BLASTN 1085 1e−81 80 2492 16 LIB3055-011-Q1-N1-G2 LIB3055 g1421750 BLASTN 958 1e−78 79 2493 16 700663981H1 SOYMON005 g1421750 BLASTN 996 1e−74 82 2494 16 701109737H1 SOYMON036 g1421750 BLASTN 955 1e−70 83 2495 16 701130190H1 SOYMON037 g1421750 BLASTN 935 1e−69 81 2496 16 701003301H1 SOYMON019 g1421750 BLASTN 412 1e−68 82 2497 16 700894137H1 SOYMON024 g1421750 BLASTN 923 1e−68 84 2498 16 700980096H1 SOYMON009 g1421750 BLASTN 923 1e−68 81 2499 16 700942625H1 SOYMON024 g1421750 BLASTN 911 1e−67 80 2500 16 700829695H1 SOYMON019 g1421750 BLASTN 918 1e−67 84 2501 16 700662572H1 SOYMON005 g1421750 BLASTN 920 1e−67 84 2502 16 701127647H1 SOYMON037 g1421750 BLASTN 902 1e−66 84 2503 16 701014550H1 SOYMON019 g1421750 BLASTN 556 1e−65 81 2504 16 LIB3051-043-Q1-K1-G3 LIB3051 g1421750 BLASTN 698 1e−65 82 2505 16 700730914H1 SOYMON009 g1421750 BLASTN 897 1e−65 84 2506 16 701052455H1 SOYMON032 g1421750 BLASTN 876 1e−64 81 2507 16 LIB3051-074-Q1-K1-A7 LIB3051 g1421750 BLASTN 876 1e−64 75 2508 16 700874839H1 SOYMON018 g1421750 BLASTN 862 1e−63 82 2509 16 701005943H1 SOYMON019 g1421750 BLASTN 865 1e−63 82 2510 16 700663357H1 SOYMON005 g1421750 BLASTN 869 1e−63 82 2511 16 701042053H1 SOYMON029 g1421750 BLASTN 872 1e−63 81 2512 16 700971988H1 SOYMON005 g1421750 BLASTN 508 1e−61 82 2513 16 701010728H1 SOYMON019 g1421750 BLASTN 657 1e−61 81 2514 16 700987206H1 SOYMON009 g1421750 BLASTN 845 1e−61 83 2515 16 701126773H1 SOYMON037 g1421750 BLASTN 847 1e−61 79 2516 16 700764714H1 SOYMON023 g1421750 BLASTN 830 1e−60 81 2517 16 700974444H1 SOYMON005 g1421750 BLASTN 830 1e−60 81 2518 16 701118620H1 SOYMON037 g1421750 BLASTN 832 1e−60 75 2519 16 700729926H1 SOYMON009 g1421750 BLASTN 834 1e−60 79 2520 16 700945142H1 SOYMON024 g1421750 BLASTN 470 1e−59 84 2521 16 700867912H1 SOYMON016 g1421750 BLASTN 820 1e−59 82 2522 16 700746546H1 SOYMON013 g1421750 BLASTN 824 1e−59 78 2523 16 700873331H1 SOYMON018 g1421750 BLASTN 769 1e−58 82 2524 16 700738212H1 SOYMON012 g1421750 BLASTN 780 1e−56 83 2525 16 700830454H1 SOYMON019 g1421750 BLASTN 789 1e−56 81 2526 16 700726129H1 SOYMON009 g1421750 BLASTN 789 1e−56 81 2527 16 700996774H1 SOYMON018 g1421750 BLASTN 486 1e−55 81 2528 16 700746427H1 SOYMON013 g1421750 BLASTN 766 1e−55 77 2529 16 701123911H1 SOYMON037 g1421750 BLASTN 769 1e−55 76 2530 16 700745730H1 SOYMON013 g1421750 BLASTN 775 1e−55 76 2531 16 700900940H1 SOYMON027 g1421750 BLASTN 760 1e−54 78 2532 16 700846419H1 SOYMON021 g1421750 BLASTN 762 1e−54 78 2533 16 701203427H1 SOYMON035 g1421750 BLASTN 491 1e−53 84 2534 16 LIB3051-074-Q1-K1-F5 LIB3051 g1421750 BLASTN 650 1e−53 75 2535 16 700747646H1 SOYMON013 g1421750 BLASTN 747 1e−53 77 2536 16 701049823H1 SOYMON032 g1421750 BLASTN 751 1e−53 77 2537 16 701049188H1 SOYMON032 g1421750 BLASTN 731 1e−52 75 2538 16 700875156H1 SOYMON018 g1421750 BLASTN 737 1e−52 78 2539 16 700864540H1 SOYMON016 g1421750 BLASTN 741 1e−52 81 2540 16 700682329H2 SOYMON008 g1421750 BLASTN 719 1e−51 76 2541 16 700846215H1 SOYMON021 g1421750 BLASTN 575 1e−50 82 2542 16 701101327H1 SOYMON028 g1421750 BLASTN 592 1e−50 77 2543 16 LIB3055-011-Q1-N1-E7 LIB3055 g1421750 BLASTN 680 1e−50 80 2544 16 700848992H1 SOYMON021 g1421750 BLASTN 708 1e−50 76 2545 16 700966820H1 SOYMON028 g1421750 BLASTN 710 1e−50 75 2546 16 701051785H1 SOYMON032 g1421750 BLASTN 717 1e−50 75 2547 16 700681079H1 SOYMON008 g1917012 BLASTN 290 1e−49 77 2548 16 701138880H1 SOYMON038 g1421750 BLASTN 610 1e−49 78 2549 16 700872967H1 SOYMON018 g1421750 BLASTN 704 1e−49 75 2550 16 701119960H1 SOYMON037 g1421750 BLASTN 684 1e−48 81 2551 16 700873306H1 SOYMON018 g1421750 BLASTN 693 1e−48 83 2552 16 700751484H1 SOYMON014 g1421750 BLASTN 635 1e−47 82 2553 16 700958865H1 SOYMON022 g1421750 BLASTN 486 1e−46 82 2554 16 700872833H1 SOYMON018 g1421750 BLASTN 659 1e−46 84 2555 16 700871673H1 SOYMON018 g1421750 BLASTN 659 1e−46 84 2556 16 700872801H1 SOYMON018 g1421750 BLASTN 659 1e−46 84 2557 16 700847572H1 SOYMON021 g1421750 BLASTN 660 1e−46 75 2558 16 700728046H1 SOYMON009 g1421750 BLASTN 661 1e−46 84 2559 16 700894721H1 SOYMON024 g1421750 BLASTN 550 1e−43 84 2560 16 700730946H1 SOYMON009 g1421750 BLASTN 307 1e−42 77 2561 16 700727943H1 SOYMON009 g1421750 BLASTN 378 1e−42 79 2562 16 701101238H1 SOYMON028 g1421750 BLASTN 456 1e−42 92 2563 16 700758519H1 SOYMON015 g1421750 BLASTN 358 1e−41 77 2564 16 700740343H1 SOYMON012 g1531764 BLASTN 607 1e−41 72 2565 16 700848076H1 SOYMON021 g1531764 BLASTN 469 1e−40 89 2566 16 LIB3051-080-Q1-K1-H7 LIB3051 g1531764 BLASTN 472 1e−38 90 2567 16 700955916H1 SOYMON022 g1531764 BLASTN 427 1e−37 90 2568 16 700873876H1 SOYMON018 g1421750 BLASTN 465 1e−37 80 2569 16 700746271H1 SOYMON013 g1421750 BLASTN 515 1e−37 80 2570 16 700662887H1 SOYMON005 g1421750 BLASTN 506 1e−35 82 2571 16 700983591H1 SOYMON009 g1421750 BLASTN 537 1e−35 82 2572 16 700752343H1 SOYMON014 g1421750 BLASTN 488 1e−34 73 2573 16 700901973H1 SOYMON027 g1421750 BLASTN 490 1e−34 82 2574 16 LIB3053-013-Q1-N1-B11 LIB3053 g1421750 BLASTN 522 1e−34 75 2575 16 700988481H1 SOYMON009 g1421750 BLASTN 506 1e−33 78 2576 16 LIB3051-078-Q1-K1-C12 LIB3051 g1421750 BLASTN 302 1e−32 75 2577 16 700985640H1 SOYMON009 g1421750 BLASTN 449 1e−30 86 2578 16 701127038H1 SOYMON037 g1421750 BLASTN 467 1e−30 67 2579 16 700898677H1 SOYMON027 g1531764 BLASTN 474 1e−30 89 2580 16 700742260H1 SOYMON012 g1421750 BLASTN 476 1e−30 70 2581 16 701103203H1 SOYMON028 g1421750 BLASTN 287 1e−27 81 2582 16 700897144H1 SOYMON027 g1421750 BLASTN 264 1e−26 75 2583 16 700832039H1 SOYMON019 g1421750 BLASTN 390 1e−26 82 2584 16 700743311H1 SOYMON012 g1421750 BLASTN 429 1e−26 87 2585 16 LIB3040-044-Q1-E1-B8 LIB3040 g1421750 BLASTN 282 1e−25 92 2586 16 LIB3051-024-Q1-K1-C4 LIB3051 g2394382 BLASTX 110 1e−24 95 2587 16 701011765H1 SOYMON019 g1421750 BLASTN 383 1e−21 76 2588 16 700893428H1 SOYMON024 g1421752 BLASTX 106 1e−20 82 2589 16 700996204H1 SOYMON018 g1421750 BLASTN 288 1e−20 75 2590 16 LIB3049-028-Q1-E1-C6 LIB3049 g1421750 BLASTN 230 1e−19 90 2591 16 701103628H1 SOYMON028 g1421750 BLASTN 277 1e−18 73 2592 16 700972636H1 SOYMON005 g1421750 BLASTN 286 1e−16 73 2593 16 700875417H1 SOYMON018 g1421750 BLASTN 209 1e−15 75 2594 16 701207702H1 SOYMON035 g1421752 BLASTX 153 1e−14 83 2595 16 701054556H1 SOYMON032 g1421750 BLASTN 230 1e−14 75 2596 16 701046407H1 SOYMON032 g1421750 BLASTN 230 1e−14 75 2597 16 701117634H1 SOYMON037 g1421750 BLASTN 230 1e−14 74 2598 16 701127714H1 SOYMON037 g1421750 BLASTN 230 1e−14 75 2599 16 700663239H1 SOYMON005 g1421750 BLASTN 210 1e−13 74 2600 16 700738510H1 SOYMON012 g1421750 BLASTN 212 1e−13 72 2601 16 700943559H1 SOYMON024 g1421750 BLASTN 215 1e−13 71 2602 16 700666278H1 SOYMON005 g1421750 BLASTN 290 1e−13 93 2603 16 700663144H1 SOYMON005 g1421750 BLASTN 204 1e−12 74 2604 16 700684147H1 SOYMON008 g1421750 BLASTN 278 1e−12 89 2605 16 700953076H1 SOYMON022 g1421750 BLASTN 285 1e−12 92 2606 16 701099931H1 SOYMON028 g1421750 BLASTN 202 1e−11 72 2607 16 701058910H1 SOYMON033 g1421750 BLASTN 216 1e−11 74 2608 16 700754949H1 SOYMON014 g1421750 BLASTN 266 1e−11 89 2609 16 700562967H1 SOYMON002 g1421750 BLASTN 266 1e−11 82 2610 16 701012790H1 SOYMON019 g1421750 BLASTN 251 1e−10 89 2611 16 701101739H1 SOYMON028 g1421750 BLASTN 260 1e−10 82 2612 16 701118211H1 SOYMON037 g1490554 BLASTX 97 1e−9 47 2613 16 700844394H1 SOYMON021 g1917013 BLASTX 116 1e−9 85 2614 16 701048560H1 SOYMON032 g1421750 BLASTN 198 1e−9 79 2615 16 701062373H1 SOYMON033 g1421750 BLASTN 203 1e−9 71 2616 16 701120496H1 SOYMON037 g1421750 BLASTN 230 1e−9 82 2617 16 701049680H1 SOYMON032 g1421750 BLASTN 177 1e−8 70 2618 16 700748760H1 SOYMON013 g1421750 BLASTN 230 1e−8 91 2619 16 700889482H1 SOYMON024 g1421750 BLASTN 235 1e−8 91 2620 16 700725013H1 SOYMON009 g1421750 BLASTN 239 1e−8 85 2621 16048 LIB3028-004-Q1-B1-E2 LIB3028 g1421750 BLASTN 879 1e−64 68 2622 16048 700761667H1 SOYMON015 g1421750 BLASTN 528 1e−35 71 2623 16048 700958003H1 SOYMON022 g1421750 BLASTN 428 1e−26 78

ASPARTATE KINASE (EC 2.7.2.4) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 858 -700018870 700018870H1 SATMON001 g500850 BLASTN 1095 1e−82 100 859 -700085903 700085903H1 SATMON011 g500852 BLASTN 1606 1e−124 99 860 -700086169 700086169H1 SATMON011 g500852 BLASTN 559 1e−94 92 861 -700096679 700096679H1 SATMON008 g500850 BLASTN 1131 1e−85 99 862 -700096794 700096794H1 SATMON008 g500850 BLASTN 1515 1e−117 100 863 -700106390 700106390H1 SATMON010 g2243115 BLASTN 544 1e−51 72 864 -700168286 700168286H1 SATMON013 g2243115 BLASTN 519 1e−34 72 865 -700171363 700171363H1 SATMON013 g500850 BLASTN 1085 1e−81 94 866 -700194781 700194781H1 SATMON014 g2257742 BLASTN 635 1e−44 79 867 -700213839 700213839H1 SATMON016 g500850 BLASTN 664 1e−75 94 868 -700219756 700219756H1 SATMON011 g500850 BLASTN 1359 1e−104 99 869 -700258808 700258808H1 SATMON017 g500850 BLASTN 630 1e−85 96 870 -700263439 700263439H1 SATMON017 g2243115 BLASTN 448 1e−43 76 871 -700266615 700266615H1 SATMON017 g2243116 BLASTX 179 1e−17 73 872 -700342655 700342655H1 SATMON021 g2257743 BLASTX 213 1e−22 82 873 -700343285 700343285H1 SATMON021 g2257742 BLASTN 437 1e−25 66 874 -700467533 700467533H1 SATMON025 g2243115 BLASTN 439 1e−26 79 875 -700548678 700548678H1 SATMON022 g147979 BLASTX 147 1e−13 62 876 -700613618 700613618H1 SATMON033 g500850 BLASTN 965 1e−81 98 877 -L30691987 LIB3069-018-Q1-K1-A3 LIB3069 g2243115 BLASTN 266 1e−10 57 878 12201 700457103H1 SATMON029 g2257742 BLASTN 789 1e−56 76 879 12201 700457111H1 SATMON029 g2243115 BLASTN 513 1e−33 76 880 12931 700380864H1 SATMON023 g500852 BLASTN 1450 1e−111 95 881 12931 700105610H1 SATMON010 g500852 BLASTN 1046 1e−105 95 882 12931 700380848H1 SATMON023 g500852 BLASTN 1193 1e−96 95 883 12931 700205392H1 SATMON003 g500852 BLASTN 1065 1e−94 93 884 12931 700552314H1 SATMON022 g500852 BLASTN 908 1e−86 90 885 12931 700551915H1 SATMON022 g500852 BLASTN 1090 1e−81 90 886 16037 700344509H1 SATMON021 g500852 BLASTN 1184 1e−89 92 887 16037 700345170H1 SATMON021 g500852 BLASTN 600 1e−58 85 888 16157 700212607H1 SATMON016 g2243115 BLASTN 914 1e−67 76 889 16157 700094809H1 SATMON008 g2243116 BLASTX 144 1e−12 84 890 19231 700091761H1 SATMON011 g500850 BLASTN 1358 1e−104 98 891 19231 700612568H1 SATMON033 g500850 BLASTN 1137 1e−102 99 892 22303 700553291H1 SATMON022 g2243115 BLASTN 644 1e−44 70 893 22303 700553382H1 SATMON022 g2243115 BLASTN 418 1e−39 70 894 28000 LIB143-061-Q1-E1-C7 LIB143 g2243115 BLASTN 1132 1e−85 75 895 28000 700474110H1 SATMON025 g2243115 BLASTN 509 1e−33 75 896 30401 700620948H1 SATMON034 g500852 BLASTN 327 1e−30 88 897 32907 LIB143-038-Q1-E1-B11 LIB143 g500850 BLASTN 1864 1e−146 96 898 32907 700096779H1 SATMON008 g500850 BLASTN 1490 1e−115 97 899 5616 700346488H1 SATMON021 g2243115 BLASTN 664 1e−46 71 900 5616 700196138H1 SATMON014 g2243115 BLASTN 630 1e−43 74 2624 -700556108 700556108H1 SOYMON001 g2243115 BLASTN 700 1e−49 74 2625 -700663367 700663367H1 SOYMON005 g2243115 BLASTN 737 1e−52 77 2626 -700733301 700733301H1 SOYMON010 g2243115 BLASTN 751 1e−53 78 2627 -700747979 700747979H1 SOYMON013 g2257742 BLASTN 449 1e−27 70 2628 -700832664 700832664H1 SOYMON019 g167547 BLASTN 322 1e−44 78 2629 -700843925 700843925H1 SOYMON021 g167547 BLASTN 616 1e−42 71 2630 -700888516 700888516H1 SOYMON024 g464225 BLASTX 193 1e−19 78 2631 -700892002 700892002H1 SOYMON024 g2243115 BLASTN 363 1e−21 79 2632 -700959057 700959057H1 SOYMON022 g2257742 BLASTN 497 1e−32 72 2633 -700971891 700971891H1 SOYMON005 g167547 BLASTN 699 1e−49 74 2634 -700984812 700984812H1 SOYMON009 g2257742 BLASTN 801 1e−57 77 2635 -701069254 701069254H1 SOYMON034 g2243115 BLASTN 260 1e−23 75 2636 -701120341 701120341H1 SOYMON037 g2243115 BLASTN 567 1e−38 77 2637 -GM35173 LIB3051-037-Q1-K1-B9 LIB3051 g2970554 BLASTN 193 1e−11 83 2638 15020 700557507H1 SOYMON001 g167547 BLASTN 914 1e−67 79 2639 15020 700666142H1 SOYMON005 g1107460 BLASTN 767 1e−55 77 2640 18237 700797368H1 SOYMON017 g2257742 BLASTN 819 1e−59 81 2641 18237 700797360H1 SOYMON017 g2257742 BLASTN 809 1e−58 84 2642 19332 LIB3056-004-Q1-N1-D5 LIB3056 g2243115 BLASTN 1118 1e−84 74 2643 19332 700786255H2 SOYMON011 g2257742 BLASTN 626 1e−43 72 2644 19332 700684751H1 SOYMON008 g2257742 BLASTN 583 1e−39 73 2645 21954 701100440H1 SOYMON028 g167547 BLASTN 835 1e−60 78 2646 21954 701059173H1 SOYMON033 g167547 BLASTN 719 1e−51 75 2647 26336 701003103H1 SOYMON019 g2243115 BLASTN 877 1e−64 79 2648 26336 700976874H1 SOYMON009 g2243115 BLASTN 880 1e−64 79

ASPARTATE-SEMIALDEHYDE DEHYDROGENASE (EC 1.2.1.11) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 901 -700439614 700439614H1 SATMON026 g2314350 BLASTX 107 1e−13 52 902 -701183695 701183695H1 SATMONN06 g289910 BLASTX 150 1e−13 69 903 -L1487398 LIB148-064-Q1-E1-D10 LIB148 g1749466 BLASTX 178 1e−33 55 904 -L30622830 LIB3062-028-Q1-K1-A8 LIB3062 g1085109 BLASTX 108 1e−40 48 2649 -700756763 700756763H1 SOYMON014 g1359593 BLASTX 71 1e−8 52 2650 -700830054 700830054H1 SOYMON019 g142828 BLASTX 87 1e−10 41 2651 -701105617 701105617H1 SOYMON036 g142828 BLASTX 215 1e−22 57 2652 -GM8539 LIB3039-047-Q1-E1-C6 LIB3039 g142828 BLASTX 188 1e−35 50 2653 30187 LIB3049-001-Q1-E1-F2 LIB3049 g1359593 BLASTX 136 1e−26 54 2654 30187 700556105H1 SOYMON001 g1359593 BLASTX 132 1e−11 56

O-SUCCINYLHOMOSERINE (THIOL)-LYASE (EC 4.2.99.9) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 905 -700049526 700049526H1 SATMON003 g2198852 BLASTN 251 1e−9 76 906 -700086788 700086788H1 SATMON011 g2198850 BLASTN 631 1e−61 88 907 -700460589 700460589H1 SATMON030 g2198850 BLASTN 251 1e−32 83 908 -700577561 700577561H1 SATMON031 g2198852 BLASTN 207 1e−15 82 909 -700579840 700579840H1 SATMON031 g2198850 BLASTN 216 1e−12 94 910 -700616013 700616013H1 SATMON033 g2198852 BLASTN 278 1e−39 81 911 -L1892785 LIB189-011-Q1-E1-A10 LIB189 g2198852 BLASTN 311 1e−14 82 912 -L30594402 LIB3059-042-Q1-K1-E11 LIB3059 g2198852 BLASTN 489 1e−40 84 913 -L30604293 LIB3060-028-Q1-K1-E7 LIB3060 g2198852 BLASTN 380 1e−20 83 914 -L30693289 LIB3069-016-Q1-K1-G10 LIB3069 g2198852 BLASTN 197 1e−10 79 915 10571 700224881H1 SATMON011 g2198852 BLASTN 319 1e−15 75 916 10801 700429049H1 SATMONN01 g2198852 BLASTN 210 1e−16 82 917 10801 700167813H1 SATMON013 g2198852 BLASTN 200 1e−15 82 918 10801 700074027H1 SATMON007 g2198852 BLASTN 178 1e−11 80 919 16379 LIB3061-035-Q1-K1-B6 LIB3061 g2198850 BLASTN 2184 1e−173 99 920 16379 700259309H1 SATMON017 g2198850 BLASTN 1259 1e−108 92 921 16379 700051696H1 SATMON003 g2198850 BLASTN 1392 1e−107 96 922 16379 700239553H1 SATMON010 g2198850 BLASTN 1265 1e−96 96 923 16379 700042846H1 SATMON004 g2198850 BLASTN 1208 1e−91 95 924 16379 700092741H1 SATMON008 g2198850 BLASTN 1042 1e−89 95 925 16379 LIB3061-017-Q1-K1-D7 LIB3061 g2198850 BLASTN 1183 1e−89 99 926 16379 700206765H1 SATMON003 g2198850 BLASTN 1095 1e−82 81 927 16379 700150281H1 SATMON007 g2198850 BLASTN 1024 1e−76 94 928 16379 700165862H1 SATMON013 g2198850 BLASTN 948 1e−75 94 929 2221 LIB3060-017-Q1-K1-B10 LIB3060 g2198850 BLASTN 1819 1e−161 99 930 2221 LIB84-006-Q1-E1-F3 LIB84 g2198852 BLASTN 1615 1e−153 97 931 2221 700575334H1 SATMON030 g2198850 BLASTN 1570 1e−124 99 932 2221 700206250H1 SATMON003 g2198850 BLASTN 1515 1e−117 100 933 2221 700095073H1 SATMON008 g2198852 BLASTN 1490 1e−115 100 934 2221 700571230H1 SATMON030 g2198850 BLASTN 1355 1e−114 93 935 2221 700157358H1 SATMON012 g2198850 BLASTN 1370 1e−105 100 936 2221 700379811H1 SATMON021 g2198850 BLASTN 1345 1e−103 93 937 2221 700041570H1 SATMON004 g2198850 BLASTN 1325 1e−101 100 938 2221 700104063H1 SATMON010 g2198850 BLASTN 1157 1e−95 90 939 2221 700378265H1 SATMON019 g2198850 BLASTN 771 1e−94 99 940 2221 700235329H1 SATMON010 g2198850 BLASTN 1234 1e−94 93 941 2221 LIB3068-057-Q1-K1-D1 LIB3068 g2198852 BLASTN 1209 1e−91 94 942 2221 700159158H1 SATMON012 g2198850 BLASTN 1174 1e−89 94 943 2221 700580854H1 SATMON031 g2198850 BLASTN 754 1e−86 92 944 2221 700623409H1 SATMON034 g2198850 BLASTN 910 1e−84 96 945 2221 701164706H1 SATMONN04 g2198852 BLASTN 577 1e−83 94 946 2221 700164719H1 SATMON013 g2198852 BLASTN 831 1e−83 99 947 2221 700158146H1 SATMON012 g2198850 BLASTN 1100 1e−82 93 948 2221 700203970H1 SATMON003 g2198852 BLASTN 996 1e−80 99 949 2221 700158313H1 SATMON012 g2198850 BLASTN 1036 1e−77 93 950 2221 700425211H1 SATMONN01 g2198852 BLASTN 485 1e−61 96 951 2221 700167764H1 SATMON013 g2198850 BLASTN 793 1e−57 93 952 23788 700102780H1 SATMON010 g2198852 BLASTN 1131 1e−104 99 953 23788 701167458H1 SATMONN05 g2198852 BLASTN 1069 1e−90 93 2655 -700900206 700900206H1 SOYMON027 g1742961 BLASTX 215 1e−24 78 2656 -GM40351 LIB3051-114-Q1-K1-H12 LIB3051 g2198851 BLASTX 122 1e−25 96 2657 12502 701101592H1 SOYMON028 g146846 BLASTX 103 1e−18 44 2658 12502 701106834H1 SOYMON036 g146846 BLASTX 103 1e−18 44 2659 13820 LIB3055-003-Q1-N1-F12 LIB3055 g3202028 BLASTX 193 1e−35 94 2660 8119 700989656H1 SOYMON011 g1742960 BLASTN 815 1e−59 79

CYSTATHIONINE β-LYASE (EC 4.4.1.8) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 954 -700155172 700155172H1 SATMON007 g704397 BLASTX 361 1e−43 78 955 -L362943 LIB36-013-Q1-E1-G10 LIB36 g704396 BLASTN 818 1e−59 75 956 19856 700240664H1 SATMON010 g704396 BLASTN 496 1e−31 69 957 19856 700572496H1 SATMON030 g704396 BLASTN 446 1e−26 68 958 22960 701170964H1 SATMONN05 g704396 BLASTN 778 1e−56 78 959 22960 LIB3061-002-Q1-K2-F9 LIB3061 g704396 BLASTN 765 1e−53 77 960 22960 701172780H2 SATMONN05 g704396 BLASTN 686 1e−48 73 961 22960 700578571H1 SATMON031 g704397 BLASTX 222 1e−23 89 962 30752 LIB3078-055-Q1-K1-C8 LIB3078 g704396 BLASTN 747 1e−51 71 963 30752 700086603H1 SATMON011 g704397 BLASTX 192 1e−18 64 2661 -701001147 701001147H1 SOYMON018 g704396 BLASTN 847 1e−61 78 2662 18602 700566066H1 SOYMON002 g704396 BLASTN 751 1e−57 79 2663 18602 700890955H1 SOYMON024 g704396 BLASTN 698 1e−49 77 2664 18602 700896865H1 SOYMON027 g704396 BLASTN 682 1e−48 77 2665 5144 LIB3050-006-Q1-E1-A9 LIB3050 g1399263 BLASTX 96 1e−31 41

5-METHYLTETRAHYDROPTEROYLTRIGLUTAMATE - HOMOCYSTEINE S-METHYLTRANSFERASE (EC 2.1.1.14) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 964 -700212217 700212217H1 SATMON016 g886471 BLASTX 151 1e−16 93 965 -700333966 700333966H1 SATMON019 g886470 BLASTN 370 1e−23 66 966 -700377403 700377403H1 SATMON019 g2738247 BLASTN 305 1e−26 80 967 -700571893 700571893H1 SATMON030 g974781 BLASTN 262 1e−20 80 968 -701165656 701165656H1 SATMONN04 g2738247 BLASTN 450 1e−50 73 969 -L30622954 LIB3062-030-Q1-K1-C9 LIB3062 g886470 BLASTN 366 1e−26 81 970 -L30662004 LIB3066-026-Q1-K1-F1 LIB3066 g2738248 BLASTX 140 1e−28 77 971 -L30671450 LIB3067-001-Q1-K1-C6 LIB3067 g2738247 BLASTN 553 1e−46 69 972 -L30694410 LIB3069-057-Q1-K1-B4 LIB3069 g886470 BLASTN 447 1e−26 67 973 1 700452404H1 SATMON028 g453939 BLASTX 59 1e−17 94 974 13513 700049012H1 SATMON003 g1814402 BLASTN 1006 1e−74 81 975 13513 700086058H1 SATMON011 g1814402 BLASTN 982 1e−72 81 976 13513 700235814H1 SATMON010 g1814402 BLASTN 890 1e−65 79 977 13513 700170015H1 SATMON013 g1814402 BLASTN 578 1e−39 78 978 3835 700093161H1 SATMON008 g974781 BLASTN 858 1e−62 76 979 3835 700223321H1 SATMON011 g974781 BLASTN 836 1e−60 80 980 3835 700454003H1 SATMON029 g974781 BLASTN 823 1e−59 82 981 3835 700238925H1 SATMON010 g2738247 BLASTN 733 1e−55 74 982 3835 700075142H1 SATMON007 g2738247 BLASTN 569 1e−52 73 983 3835 700151969H1 SATMON007 g974781 BLASTN 660 1e−46 76 984 3835 700281434H2 SATMON019 g974781 BLASTN 560 1e−45 79 985 3835 700084914H1 SATMON011 g974781 BLASTN 440 1e−36 79 986 3835 700215135H1 SATMON016 g974781 BLASTN 543 1e−36 78 987 3835 700202218H1 SATMON003 g974781 BLASTN 389 1e−23 78 988 3835 700281467H2 SATMON019 g2738248 BLASTX 123 1e−12 71 989 456 LIB3059-019-Q1-K1-G6 LIB3059 g974781 BLASTN 1493 1e−115 83 990 456 LIB3068-012-Q1-K1-G7 LIB3068 g974781 BLASTN 1471 1e−113 81 991 456 LIB3061-050-Q1-K1-G10 LIB3061 g886470 BLASTN 1285 1e−98 82 992 456 LIB3067-043-Q1-K1-F9 LIB3067 g2738247 BLASTN 1287 1e−98 79 993 456 700087169H1 SATMON011 g1814402 BLASTN 1245 1e−94 86 994 456 LIB3069-030-Q1-K1-G9 LIB3069 g1814402 BLASTN 772 1e−93 80 995 456 LIB3069-019-Q1-K1-G11 LIB3069 g1814402 BLASTN 1202 1e−91 81 996 456 700202514H1 SATMON003 g1814402 BLASTN 1134 1e−89 84 997 456 LIB3062-011-Q1-K1-F2 LIB3062 g886470 BLASTN 664 1e−86 84 998 456 LIB143-015-Q1-E1-A1 LIB143 g974781 BLASTN 1143 1e−86 81 999 456 700570326H1 SATMON030 g974781 BLASTN 1089 1e−85 83 1000 456 700103727H1 SATMON010 g2738247 BLASTN 1132 1e−85 86 1001 456 700574823H1 SATMON030 g1814402 BLASTN 1135 1e−85 80 1002 456 700091666H1 SATMON011 g1814402 BLASTN 1136 1e−85 84 1003 456 LIB3069-033-Q1-K1-E9 LIB3069 g2738247 BLASTN 1115 1e−84 79 1004 456 700572634H1 SATMON030 g1814402 BLASTN 1123 1e−84 85 1005 456 700211829H1 SATMON016 g2738247 BLASTN 1096 1e−82 84 1006 456 700087109H1 SATMON011 g1814402 BLASTN 1102 1e−82 85 1007 456 LIB3059-003-Q1-K1-B1 LIB3059 g1814402 BLASTN 961 1e−81 82 1008 456 700047556H1 SATMON003 g2738247 BLASTN 976 1e−81 82 1009 456 700201925H1 SATMON003 g2738247 BLASTN 1001 1e−81 84 1010 456 700209343H1 SATMON016 g1814402 BLASTN 1081 1e−81 82 1011 456 700348416H1 SATMON023 g1814402 BLASTN 1081 1e−81 83 1012 456 700095559H1 SATMON008 g1814402 BLASTN 1085 1e−81 82 1013 456 700073472H1 SATMON007 g1814402 BLASTN 1089 1e−81 82 1014 456 LIB3062-057-Q1-K1-C2 LIB3062 g886470 BLASTN 1073 1e−80 78 1015 456 700093445H1 SATMON008 g1814402 BLASTN 1076 1e−80 84 1016 456 LIB143-026-Q1-E1-H3 LIB143 g2738247 BLASTN 648 1e−79 79 1017 456 700206240H1 SATMON003 g1814402 BLASTN 1057 1e−79 82 1018 456 700091907H1 SATMON011 g886470 BLASTN 1062 1e−79 80 1019 456 700258178H1 SATMON017 g1814402 BLASTN 1064 1e−79 84 1020 456 700086565H1 SATMON011 g1814402 BLASTN 1065 1e−79 82 1021 456 700471452H1 SATMON025 g2738247 BLASTN 1045 1e−78 85 1022 456 700243858H1 SATMON010 g1814402 BLASTN 1048 1e−78 85 1023 456 700352224H1 SATMON023 g2738247 BLASTN 1051 1e−78 84 1024 456 700084769H1 SATMON011 g1814402 BLASTN 1054 1e−78 83 1025 456 700082818H1 SATMON011 g886470 BLASTN 984 1e−77 84 1026 456 700331974H1 SATMON019 g886470 BLASTN 1031 1e−77 82 1027 456 700086203H1 SATMON011 g1814402 BLASTN 1032 1e−77 81 1028 456 700075826H1 SATMON007 g2738247 BLASTN 1038 1e−77 82 1029 456 700104555H1 SATMON010 g886470 BLASTN 1039 1e−77 79 1030 456 700074084H1 SATMON007 g2738247 BLASTN 1039 1e−77 87 1031 456 700076872H1 SATMON007 g1814402 BLASTN 700 1e−76 83 1032 456 700050704H1 SATMON003 g1814402 BLASTN 880 1e−76 82 1033 456 700030215H1 SATMON003 g974781 BLASTN 1019 1e−76 83 1034 456 700077416H1 SATMON007 g886470 BLASTN 1019 1e−76 79 1035 456 700093982H1 SATMON008 g1814402 BLASTN 1019 1e−76 82 1036 456 700048847H1 SATMON003 g2738247 BLASTN 1023 1e−76 85 1037 456 700090739H1 SATMON011 g886470 BLASTN 1026 1e−76 82 1038 456 700092904H1 SATMON008 g886470 BLASTN 1026 1e−76 82 1039 456 LIB3068-009-Q1-K1-E5 LIB3068 g1814402 BLASTN 1027 1e−76 73 1040 456 700336705H1 SATMON019 g1814402 BLASTN 747 1e−75 83 1041 456 700344872H1 SATMON021 g1814402 BLASTN 768 1e−75 85 1042 456 LIB3066-019-Q1-K1-E1 LIB3066 g886470 BLASTN 868 1e−75 81 1043 456 700086322H1 SATMON011 g2738247 BLASTN 1015 1e−75 81 1044 456 LIB3067-052-Q1-K1-B12 LIB3067 g974781 BLASTN 1033 1e−75 81 1045 456 700221395H1 SATMON011 g2738247 BLASTN 1000 1e−74 85 1046 456 700092483H1 SATMON008 g1814402 BLASTN 1001 1e−74 82 1047 456 700618708H1 SATMON034 g886470 BLASTN 1003 1e−74 82 1048 456 700074191H1 SATMON007 g2738247 BLASTN 1005 1e−74 86 1049 456 LIB143-055-Q1-E1-F10 LIB143 g1814402 BLASTN 986 1e−73 83 1050 456 700025955H1 SATMON003 g1814402 BLASTN 993 1e−73 84 1051 456 700574824H1 SATMON030 g886470 BLASTN 564 1e−72 80 1052 456 700201527H1 SATMON003 g974781 BLASTN 812 1e−72 81 1053 456 700092202H1 SATMON008 g886470 BLASTN 946 1e−72 79 1054 456 700223674H1 SATMON011 g2738247 BLASTN 971 1e−72 83 1055 456 700092638H1 SATMON008 g1814402 BLASTN 972 1e−72 84 1056 456 700090025H1 SATMON011 g974781 BLASTN 973 1e−72 81 1057 456 700217069H1 SATMON016 g2738247 BLASTN 976 1e−72 83 1058 456 700349142H1 SATMON023 g1814402 BLASTN 980 1e−72 82 1059 456 700088387H1 SATMON011 g2738247 BLASTN 980 1e−72 85 1060 456 700215102H1 SATMON016 g886470 BLASTN 980 1e−72 79 1061 456 700456708H1 SATMON029 g2738247 BLASTN 982 1e−72 82 1062 456 700210025H1 SATMON016 g1814402 BLASTN 982 1e−72 81 1063 456 700335345H1 SATMON019 g974781 BLASTN 982 1e−72 84 1064 456 700085257H1 SATMON011 g1814402 BLASTN 554 1e−71 82 1065 456 700026781H1 SATMON003 g974781 BLASTN 961 1e−71 82 1066 456 700224265H1 SATMON011 g2738247 BLASTN 962 1e−71 83 1067 456 700083282H1 SATMON011 g1814402 BLASTN 962 1e−71 83 1068 456 700381324H1 SATMON023 g1814402 BLASTN 965 1e−71 82 1069 456 700212017H1 SATMON016 g1814402 BLASTN 640 1e−70 83 1070 456 700088053H1 SATMON011 g886470 BLASTN 853 1e−70 84 1071 456 LIB3062-032-Q1-K1-C2 LIB3062 g1814402 BLASTN 883 1e−70 80 1072 456 700083030H1 SATMON011 g1814402 BLASTN 948 1e−70 81 1073 456 700613980H1 SATMON033 g1814402 BLASTN 950 1e−70 83 1074 456 700347019H1 SATMON021 g974781 BLASTN 952 1e−70 81 1075 456 700335621H1 SATMON019 g1814402 BLASTN 952 1e−70 80 1076 456 700094101H1 SATMON008 g1814402 BLASTN 953 1e−70 83 1077 456 700073128H1 SATMON007 g2738247 BLASTN 954 1e−70 78 1078 456 700076245H1 SATMON007 g1814402 BLASTN 955 1e−70 83 1079 456 700071903H1 SATMON007 g886470 BLASTN 956 1e−70 81 1080 456 700090035H1 SATMON011 g886470 BLASTN 935 1e−69 80 1081 456 700405323H1 SATMON029 g1814402 BLASTN 937 1e−69 83 1082 456 700050121H1 SATMON003 g886470 BLASTN 938 1e−69 83 1083 456 700224714H1 SATMON011 g1814402 BLASTN 939 1e−69 82 1084 456 700281535H2 SATMON019 g1814402 BLASTN 940 1e−69 81 1085 456 700222093H1 SATMON011 g2738247 BLASTN 941 1e−69 81 1086 456 700094659H1 SATMON008 g1814402 BLASTN 942 1e−69 84 1087 456 700237187H1 SATMON010 g1814402 BLASTN 942 1e−69 84 1088 456 700023210H1 SATMON003 g1814402 BLASTN 943 1e−69 83 1089 456 700072813H1 SATMON007 g1814402 BLASTN 944 1e−69 82 1090 456 700075720H1 SATMON007 g1814402 BLASTN 946 1e−69 83 1091 456 700072107H1 SATMON007 g1814402 BLASTN 946 1e−69 80 1092 456 700082225H1 SATMON011 g1814402 BLASTN 691 1e−68 82 1093 456 700085411H1 SATMON011 g1814402 BLASTN 723 1e−68 83 1094 456 700454357H1 SATMON029 g2738247 BLASTN 926 1e−68 82 1095 456 700620976H1 SATMON034 g1814402 BLASTN 927 1e−68 81 1096 456 700241483H1 SATMON010 g1814402 BLASTN 931 1e−68 86 1097 456 700355459H1 SATMON024 g974781 BLASTN 931 1e−68 82 1098 456 700095875H1 SATMON008 g1814402 BLASTN 931 1e−68 87 1099 456 700076702H1 SATMON007 g1814402 BLASTN 934 1e−68 84 1100 456 700212538H1 SATMON016 g1814402 BLASTN 689 1e−67 85 1101 456 700086479H1 SATMON011 g1814402 BLASTN 715 1e−67 82 1102 456 700453684H1 SATMON028 g1814402 BLASTN 747 1e−67 83 1103 456 700204454H1 SATMON003 g1814402 BLASTN 797 1e−67 82 1104 456 700612577H1 SATMON033 g974781 BLASTN 911 1e−67 81 1105 456 700474018H1 SATMON025 g886470 BLASTN 917 1e−67 81 1106 456 700213602H1 SATMON016 g886470 BLASTN 917 1e−67 76 1107 456 700076911H1 SATMON007 g974781 BLASTN 917 1e−67 81 1108 456 700223313H1 SATMON011 g886470 BLASTN 919 1e−67 84 1109 456 700220692H1 SATMON011 g974781 BLASTN 920 1e−67 81 1110 456 700258038H1 SATMON017 g886470 BLASTN 920 1e−67 83 1111 456 700095862H1 SATMON008 g1814402 BLASTN 922 1e−67 86 1112 456 700213396H1 SATMON016 g886470 BLASTN 922 1e−67 83 1113 456 700354259H1 SATMON024 g974781 BLASTN 638 1e−66 81 1114 456 700471505H1 SATMON025 g886470 BLASTN 681 1e−66 83 1115 456 700202439H1 SATMON003 g1814402 BLASTN 706 1e−66 82 1116 456 700223609H1 SATMON011 g974781 BLASTN 770 1e−66 83 1117 456 700405260H1 SATMON028 g1814402 BLASTN 782 1e−66 86 1118 456 700085214H1 SATMON011 g886470 BLASTN 814 1e−66 84 1119 456 700211194H1 SATMON016 g1814402 BLASTN 902 1e−66 84 1120 456 700106641H1 SATMON010 g1814402 BLASTN 903 1e−66 82 1121 456 700405170H1 SATMON028 g974781 BLASTN 904 1e−66 79 1122 456 700090015H1 SATMON011 g886470 BLASTN 905 1e−66 80 1123 456 700623154H1 SATMON034 g886470 BLASTN 906 1e−66 81 1124 456 700458895H1 SATMON029 g886470 BLASTN 906 1e−66 83 1125 456 700094073H1 SATMON008 g886470 BLASTN 907 1e−66 78 1126 456 700158508H1 SATMON012 g2738247 BLASTN 909 1e−66 82 1127 456 700027366H1 SATMON003 g1814402 BLASTN 910 1e−66 81 1128 456 700073636H1 SATMON007 g1814402 BLASTN 615 1e−65 87 1129 456 700220207H1 SATMON011 g886470 BLASTN 785 1e−65 84 1130 456 LIB143-007-Q1-E1-A6 LIB143 g886470 BLASTN 807 1e−65 80 1131 456 700575288H1 SATMON030 g1814402 BLASTN 887 1e−65 83 1132 456 700219870H1 SATMON011 g974781 BLASTN 887 1e−65 82 1133 456 701184573H1 SATMONN06 g974781 BLASTN 888 1e−65 81 1134 456 700076946H1 SATMON007 g1814402 BLASTN 891 1e−65 86 1135 456 700214808H1 SATMON016 g2738247 BLASTN 893 1e−65 81 1136 456 700096274H1 SATMON008 g1814402 BLASTN 894 1e−65 80 1137 456 700160445H1 SATMON012 g1814402 BLASTN 896 1e−65 85 1138 456 700071727H1 SATMON007 g974781 BLASTN 898 1e−65 80 1139 456 700571751H1 SATMON030 g1814402 BLASTN 898 1e−65 78 1140 456 700215107H1 SATMON016 g2738247 BLASTN 507 1e−64 77 1141 456 LIB3067-057-Q1-K1-C6 LIB3067 g2738247 BLASTN 555 1e−64 76 1142 456 700614743H1 SATMON033 g886470 BLASTN 748 1e−64 81 1143 456 700444971H1 SATMON027 g974781 BLASTN 803 1e−64 83 1144 456 700224573H1 SATMON011 g1814402 BLASTN 805 1e−64 86 1145 456 700019276H1 SATMON001 g1814402 BLASTN 875 1e−64 83 1146 456 700210440H1 SATMON016 g2738247 BLASTN 881 1e−64 80 1147 456 700093168H1 SATMON008 g1814402 BLASTN 885 1e−64 86 1148 456 700224478H1 SATMON011 g886470 BLASTN 886 1e−64 81 1149 456 700261958H1 SATMON017 g1814402 BLASTN 781 1e−63 79 1150 456 700074221H1 SATMON007 g1814402 BLASTN 825 1e−63 81 1151 456 700089966H1 SATMON011 g1814402 BLASTN 863 1e−63 84 1152 456 700210422H1 SATMON016 g1814402 BLASTN 863 1e−63 81 1153 456 700160272H1 SATMON012 g1814402 BLASTN 863 1e−63 85 1154 456 700444282H1 SATMON027 g2738247 BLASTN 549 1e−62 84 1155 456 700206489H1 SATMON003 g2738247 BLASTN 597 1e−62 80 1156 456 700446452H1 SATMON027 g974781 BLASTN 711 1e−62 80 1157 456 700265473H1 SATMON017 g1814402 BLASTN 715 1e−62 79 1158 456 700473708H1 SATMON025 g1814402 BLASTN 762 1e−62 82 1159 456 LIB3069-025-Q1-K1-A7 LIB3069 g886470 BLASTN 852 1e−62 80 1160 456 700166631H1 SATMON013 g2738247 BLASTN 858 1e−62 85 1161 456 700582413H1 SATMON031 g1814402 BLASTN 861 1e−62 78 1162 456 700071904H1 SATMON007 g2738247 BLASTN 544 1e−61 77 1163 456 700622655H1 SATMON034 g1814402 BLASTN 592 1e−61 82 1164 456 700082541H1 SATMON011 g1814402 BLASTN 793 1e−61 82 1165 456 700335378H1 SATMON019 g2738247 BLASTN 841 1e−61 76 1166 456 700203585H1 SATMON003 g2738247 BLASTN 846 1e−61 86 1167 456 700053043H1 SATMON007 g2738247 BLASTN 847 1e−61 81 1168 456 700163654H1 SATMON013 g1814402 BLASTN 474 1e−60 84 1169 456 700352269H1 SATMON023 g1814402 BLASTN 479 1e−60 84 1170 456 700106187H1 SATMON010 g1814402 BLASTN 631 1e−60 81 1171 456 LIB143-007-Q1-E1-A7 LIB143 g886470 BLASTN 759 1e−60 76 1172 456 700458694H1 SATMON029 g1814402 BLASTN 827 1e−60 86 1173 456 700213509H1 SATMON016 g886470 BLASTN 834 1e−60 81 1174 456 700150191H1 SATMON007 g2738247 BLASTN 834 1e−60 82 1175 456 700157594H1 SATMON012 g1814402 BLASTN 835 1e−60 80 1176 456 700094046H1 SATMON008 g974781 BLASTN 836 1e−60 79 1177 456 700220254H1 SATMON011 g2738247 BLASTN 687 1e−59 77 1178 456 700095485H1 SATMON008 g886470 BLASTN 819 1e−59 77 1179 456 LIB143-006-Q1-E1-B3 LIB143 g1814402 BLASTN 830 1e−59 65 1180 456 700071763H1 SATMON007 g2738247 BLASTN 565 1e−58 75 1181 456 700082259H1 SATMON011 g1814402 BLASTN 707 1e−58 84 1182 456 700020717H1 SATMON001 g886470 BLASTN 805 1e−58 81 1183 456 700041892H1 SATMON004 g1814402 BLASTN 805 1e−58 86 1184 456 700356573H1 SATMON024 g1814402 BLASTN 811 1e−58 81 1185 456 700171632H1 SATMON013 g2738247 BLASTN 811 1e−58 85 1186 456 700222516H1 SATMON011 g1814402 BLASTN 812 1e−58 87 1187 456 700466679H1 SATMON025 g1814402 BLASTN 812 1e−58 87 1188 456 700208929H1 SATMON016 g1814402 BLASTN 407 1e−57 78 1189 456 700355014H1 SATMON024 g1814402 BLASTN 592 1e−57 85 1190 456 701159834H1 SATMONN04 g1814402 BLASTN 683 1e−57 77 1191 456 700351137H1 SATMON023 g974781 BLASTN 725 1e−57 79 1192 456 700027308H1 SATMON003 g2738247 BLASTN 791 1e−57 80 1193 456 700142587H1 SATMON012 g974781 BLASTN 791 1e−57 83 1194 456 700170157H1 SATMON013 g1814402 BLASTN 791 1e−57 81 1195 456 700019438H1 SATMON001 g974781 BLASTN 798 1e−57 82 1196 456 700166430H1 SATMON013 g886470 BLASTN 799 1e−57 84 1197 456 700047369H1 SATMON003 g886470 BLASTN 799 1e−57 80 1198 456 700152373H1 SATMON007 g2738247 BLASTN 799 1e−57 84 1199 456 700050824H1 SATMON003 g2738247 BLASTN 605 1e−56 81 1200 456 700204588H1 SATMON003 g886470 BLASTN 720 1e−56 82 1201 456 700071667H1 SATMON007 g1814402 BLASTN 787 1e−56 85 1202 456 701185269H1 SATMONN06 g886470 BLASTN 598 1e−55 76 1203 456 700243844H1 SATMON010 g2738247 BLASTN 769 1e−55 79 1204 456 700221969H1 SATMON011 g974781 BLASTN 771 1e−55 79 1205 456 700335706H1 SATMON019 g1814402 BLASTN 773 1e−55 80 1206 456 700622285H1 SATMON034 g1814402 BLASTN 774 1e−55 82 1207 456 700089665H1 SATMON011 g2738247 BLASTN 774 1e−55 78 1208 456 700464833H1 SATMON025 g2738247 BLASTN 455 1e−54 72 1209 456 LIB3069-057-Q1-K1-A6 LIB3069 g974781 BLASTN 564 1e−54 76 1210 456 700156818H1 SATMON012 g974781 BLASTN 757 1e−54 81 1211 456 700219420H1 SATMON011 g974781 BLASTN 758 1e−54 79 1212 456 700152570H1 SATMON007 g1814402 BLASTN 761 1e−54 86 1213 456 700084484H1 SATMON011 g1814402 BLASTN 764 1e−54 84 1214 456 700152208H1 SATMON007 g1814402 BLASTN 765 1e−54 82 1215 456 700550889H1 SATMON022 g2738247 BLASTN 546 1e−53 76 1216 456 700025869H1 SATMON003 g2738247 BLASTN 675 1e−53 79 1217 456 700383075H1 SATMON024 g974781 BLASTN 724 1e−53 81 1218 456 700575104H1 SATMON030 g1814402 BLASTN 745 1e−53 70 1219 456 LIB3069-022-Q1-K1-C1 LIB3069 g974781 BLASTN 768 1e−53 81 1220 456 700215272H1 SATMON016 g1814402 BLASTN 674 1e−52 83 1221 456 700167881H1 SATMON013 g974781 BLASTN 737 1e−52 79 1222 456 700019727H1 SATMON001 g2738247 BLASTN 737 1e−52 81 1223 456 700351776H1 SATMON023 g886470 BLASTN 741 1e−52 76 1224 456 700210543H1 SATMON016 g974781 BLASTN 742 1e−52 85 1225 456 700571604H1 SATMON030 g1814402 BLASTN 360 1e−51 76 1226 456 700169777H1 SATMON013 g1814402 BLASTN 426 1e−51 83 1227 456 700096744H1 SATMON008 g2738247 BLASTN 539 1e−51 79 1228 456 700383157H1 SATMON024 g974781 BLASTN 667 1e−51 79 1229 456 700155764H1 SATMON007 g886470 BLASTN 721 1e−51 75 1230 456 700150817H1 SATMON007 g1814402 BLASTN 727 1e−51 75 1231 456 700619660H1 SATMON034 g886470 BLASTN 727 1e−51 73 1232 456 700471085H1 SATMON025 g1814402 BLASTN 406 1e−50 79 1233 456 700163226H1 SATMON013 g2738247 BLASTN 475 1e−50 78 1234 456 700444567H1 SATMON027 g1814402 BLASTN 498 1e−50 81 1235 456 700457303H1 SATMON029 g2738247 BLASTN 662 1e−50 80 1236 456 700088065H1 SATMON011 g886470 BLASTN 707 1e−50 79 1237 456 700152592H1 SATMON007 g886470 BLASTN 716 1e−50 80 1238 456 LIB3068-061-Q1-K1-B2 LIB3068 g886470 BLASTN 730 1e−50 78 1239 456 700331880H1 SATMON019 g1814402 BLASTN 697 1e−49 85 1240 456 700218025H1 SATMON016 g886470 BLASTN 700 1e−49 79 1241 456 700348643H1 SATMON023 g1814402 BLASTN 701 1e−49 80 1242 456 700236975H1 SATMON010 g974781 BLASTN 704 1e−49 81 1243 456 700442968H1 SATMON026 g886470 BLASTN 418 1e−48 79 1244 456 700479529H1 SATMON034 g2738247 BLASTN 508 1e−48 75 1245 456 700160983H1 SATMON012 g1814402 BLASTN 690 1e−48 83 1246 456 700050185H1 SATMON003 g974781 BLASTN 421 1e−47 75 1247 456 700611764H1 SATMON022 g1814402 BLASTN 503 1e−47 79 1248 456 700622669H1 SATMON034 g1814402 BLASTN 549 1e−47 79 1249 456 700153237H1 SATMON007 g1814402 BLASTN 672 1e−47 80 1250 456 700242703H1 SATMON010 g886470 BLASTN 682 1e−47 81 1251 456 700165177H1 SATMON013 g2738247 BLASTN 682 1e−47 78 1252 456 700379889H1 SATMON021 g886470 BLASTN 682 1e−47 85 1253 456 700449243H1 SATMON028 g1814402 BLASTN 357 1e−46 83 1254 456 700453282H1 SATMON028 g886470 BLASTN 608 1e−46 81 1255 456 700224308H1 SATMON011 g1814402 BLASTN 661 1e−46 83 1256 456 700150484H1 SATMON007 g886470 BLASTN 663 1e−46 78 1257 456 700240609H1 SATMON010 g2738247 BLASTN 668 1e−46 79 1258 456 700171610H1 SATMON013 g974781 BLASTN 669 1e−46 79 1259 456 LIB143-027-Q1-E1-H3 LIB143 g1814402 BLASTN 670 1e−46 86 1260 456 700334084H1 SATMON019 g1814402 BLASTN 467 1e−45 81 1261 456 LIB3069-043-Q1-K1-H6 LIB3069 g886470 BLASTN 546 1e−45 83 1262 456 700222515H1 SATMON011 g1814402 BLASTN 648 1e−45 75 1263 456 700223749H1 SATMON011 g1814402 BLASTN 648 1e−45 75 1264 456 701180187H1 SATMONN05 g974781 BLASTN 648 1e−45 78 1265 456 700050111H1 SATMON003 g2738247 BLASTN 653 1e−45 82 1266 456 700051275H1 SATMON003 g1814402 BLASTN 379 1e−44 75 1267 456 700235202H1 SATMON010 g2738247 BLASTN 637 1e−44 80 1268 456 700257385H1 SATMON017 g1814402 BLASTN 638 1e−44 80 1269 456 LIB143-028-Q1-E1-H7 LIB143 g1814402 BLASTN 638 1e−44 80 1270 456 700576066H1 SATMON030 g886470 BLASTN 516 1e−43 76 1271 456 700455065H1 SATMON029 g886470 BLASTN 624 1e−43 79 1272 456 700074158H1 SATMON007 g1814402 BLASTN 630 1e−43 83 1273 456 700171322H1 SATMON013 g1814402 BLASTN 634 1e−43 77 1274 456 700457369H1 SATMON029 g974781 BLASTN 392 1e−42 83 1275 456 700378252H1 SATMON019 g2738247 BLASTN 462 1e−42 77 1276 456 700454626H1 SATMON029 g1814402 BLASTN 495 1e−42 85 1277 456 700208182H1 SATMON016 g2738247 BLASTN 564 1e−42 73 1278 456 700618848H1 SATMON034 g886470 BLASTN 615 1e−42 80 1279 456 700222969H1 SATMON011 g1814402 BLASTN 622 1e−42 74 1280 456 700444782H1 SATMON027 g1814402 BLASTN 337 1e−41 81 1281 456 700204404H1 SATMON003 g2738247 BLASTN 607 1e−41 80 1282 456 700347028H1 SATMON021 g1814402 BLASTN 598 1e−40 81 1283 456 700172447H1 SATMON013 g1814402 BLASTN 328 1e−39 85 1284 456 700549696H1 SATMON022 g886470 BLASTN 360 1e−39 81 1285 456 700257287H1 SATMON017 g1814402 BLASTN 365 1e−38 84 1286 456 700569630H1 SATMON030 g1814402 BLASTN 566 1e−38 78 1287 456 700207258H1 SATMON017 g1814402 BLASTN 568 1e−38 85 1288 456 700052467H1 SATMON003 g1814402 BLASTN 515 1e−37 82 1289 456 700083656H1 SATMON011 g2738247 BLASTN 552 1e−37 83 1290 456 700429279H1 SATMONN01 g2738247 BLASTN 556 1e−37 79 1291 456 700349921H1 SATMON023 g2738247 BLASTN 471 1e−36 78 1292 456 700449609H1 SATMON028 g886470 BLASTN 540 1e−36 81 1293 456 700150152H1 SATMON007 g1814402 BLASTN 543 1e−36 74 1294 456 700075675H1 SATMON007 g1814402 BLASTN 546 1e−36 81 1295 456 700267015H1 SATMON017 g886470 BLASTN 550 1e−36 75 1296 456 700465118H1 SATMON025 g1814402 BLASTN 555 1e−36 78 1297 456 700456662H1 SATMON029 g2738247 BLASTN 314 1e−34 76 1298 456 700405315H1 SATMON029 g886470 BLASTN 519 1e−34 83 1299 456 700218639H1 SATMON011 g974781 BLASTN 524 1e−34 81 1300 456 700216606H1 SATMON016 g1814402 BLASTN 524 1e−34 83 1301 456 700456965H1 SATMON029 g2738247 BLASTN 525 1e−34 78 1302 456 700102147H1 SATMON010 g974781 BLASTN 526 1e−34 82 1303 456 700235734H1 SATMON010 g2738247 BLASTN 526 1e−34 83 1304 456 LIB3068-009-Q1-K1-E10 LIB3068 g886470 BLASTN 548 1e−34 70 1305 456 700029363H1 SATMON003 g974781 BLASTN 377 1e−33 80 1306 456 LIB3069-030-Q1-K1-D10 LIB3069 g886470 BLASTN 297 1e−30 78 1307 456 700334895H1 SATMON019 g974781 BLASTN 468 1e−30 82 1308 456 700150963H1 SATMON007 g974781 BLASTN 459 1e−29 88 1309 456 700152817H1 SATMON007 g974781 BLASTN 459 1e−29 88 1310 456 700208732H1 SATMON016 g974781 BLASTN 485 1e−29 76 1311 456 700051461H1 SATMON003 g886470 BLASTN 467 1e−28 73 1312 456 700616589H1 SATMON033 g2738247 BLASTN 273 1e−27 78 1313 456 700236150H1 SATMON010 g974781 BLASTN 434 1e−27 84 1314 456 700453369H1 SATMON028 g1814402 BLASTN 436 1e−27 84 1315 456 700621739H1 SATMON034 g1814402 BLASTN 436 1e−27 69 1316 456 700075162H1 SATMON007 g1814402 BLASTN 438 1e−27 85 1317 456 700405040H1 SATMON027 g974781 BLASTN 438 1e−27 72 1318 456 700356893H1 SATMON024 g974781 BLASTN 458 1e−27 82 1319 456 701185493H1 SATMONN06 g886471 BLASTX 72 1e−26 97 1320 456 700206095H1 SATMON003 g974781 BLASTN 421 1e−26 83 1321 456 700083901H1 SATMON011 g1814402 BLASTN 421 1e−26 80 1322 456 LIB3062-026-Q1-K1-D7 LIB3062 g1814402 BLASTN 423 1e−24 81 1323 456 700377277H1 SATMON019 g1814402 BLASTN 357 1e−20 75 1324 456 700201214H1 SATMON003 g2738248 BLASTX 152 1e−18 77 1325 456 700025959H1 SATMON003 g886471 BLASTX 188 1e−18 94 1326 456 700259484H1 SATMON017 g1814402 BLASTN 189 1e−17 82 1327 456 700613969H1 SATMON033 g886470 BLASTN 315 1e−17 90 1328 456 700215602H1 SATMON016 g2738248 BLASTX 158 1e−16 74 1329 456 700438152H1 SATMON026 g2738247 BLASTN 258 1e−16 75 1330 456 700458654H1 SATMON029 g886470 BLASTN 330 1e−16 70 1331 456 700449563H1 SATMON028 g886471 BLASTX 163 1e−15 89 1332 456 700236490H1 SATMON010 g1814402 BLASTN 276 1e−14 89 1333 456 700456927H1 SATMON029 g1814403 BLASTX 119 1e−13 86 1334 456 701180088H1 SATMONN05 g886471 BLASTX 152 1e−13 60 1335 456 700455716H1 SATMON029 g2738248 BLASTX 111 1e−12 67 1336 456 700573520H1 SATMON030 g886471 BLASTX 82 1e−11 84 1337 456 700569938H1 SATMON030 g2738248 BLASTX 132 1e−11 91 1338 456 700551958H1 SATMON022 g2738247 BLASTN 264 1e−11 82 1339 456 700337475H1 SATMON020 g2738247 BLASTN 268 1e−11 83 1340 456 700170827H1 SATMON013 g886471 BLASTX 124 1e−10 77 1341 456 700453382H1 SATMON028 g974781 BLASTN 152 1e−10 85 1342 456 700089211H1 SATMON011 g2738247 BLASTN 245 1e−9 79 1343 456 700103155H1 SATMON010 g2738248 BLASTX 82 1e−8 85 1344 5523 LIB3062-017-Q1-K1-F4 LIB3062 g2738247 BLASTN 932 1e−84 76 1345 5523 700210708H1 SATMON016 g2738247 BLASTN 952 1e−70 79 1346 5523 700219307H1 SATMON011 g1814402 BLASTN 892 1e−65 78 1347 5523 700221188H1 SATMON011 g2738247 BLASTN 867 1e−63 81 1348 5523 700203884H1 SATMON003 g2738247 BLASTN 874 1e−63 77 1349 5523 700218549H1 SATMON011 g2738247 BLASTN 811 1e−58 77 1350 5523 700572845H2 SATMON030 g2738247 BLASTN 785 1e−56 77 1351 5523 700152362H1 SATMON007 g886470 BLASTN 742 1e−52 79 1352 5523 700152138H1 SATMON007 g2738247 BLASTN 714 1e−50 80 1353 5523 700218842H1 SATMON011 g2738247 BLASTN 700 1e−49 75 2666 -700697958 700697958H1 SOYMON015 g2738247 BLASTN 258 1e−10 83 2667 -700731277 700731277H1 SOYMON009 g886470 BLASTN 930 1e−68 83 2668 -700749261 700749261H1 SOYMON013 g1814402 BLASTN 508 1e−38 78 2669 -700787742 700787742H2 SOYMON011 g1814402 BLASTN 648 1e−50 81 2670 -700831146 700831146H1 SOYMON019 g1814402 BLASTN 476 1e−34 80 2671 -700832029 700832029H1 SOYMON019 g1814402 BLASTN 328 1e−27 78 2672 -700854481 700854481H1 SOYMON023 g1814402 BLASTN 193 1e−11 85 2673 -700873716 700873716H1 SOYMON018 g1749542 BLASTX 99 1e−15 53 2674 -700893904 700893904H1 SOYMON024 g886470 BLASTN 413 1e−25 82 2675 -700909658 700909658H1 SOYMON022 g1814402 BLASTN 251 1e−10 91 2676 -700943841 700943841H1 SOYMON024 g886470 BLASTN 549 1e−49 82 2677 -700963223 700963223H1 SOYMON022 g886470 BLASTN 649 1e−45 72 2678 -700974103 700974103H1 SOYMON005 g886470 BLASTN 763 1e−54 80 2679 -700994293 700994293H1 SOYMON011 g974782 BLASTX 97 1e−12 72 2680 -701004816 701004816H1 SOYMON019 g1814402 BLASTN 582 1e−39 84 2681 -701007125 701007125H1 SOYMON019 g1814403 BLASTX 152 1e−13 87 2682 -701008540 701008540H1 SOYMON019 g886470 BLASTN 770 1e−55 74 2683 -701037766 701037766H1 SOYMON029 g1814403 BLASTX 72 1e−10 69 2684 -701062191 701062191H1 SOYMON033 g974781 BLASTN 225 1e−9 79 2685 -701105474 701105474H1 SOYMON036 g974782 BLASTX 164 1e−15 91 2686 -GM14442 LIB3049-056-Q1-E1-B1 LIB3049 g974781 BLASTN 231 1e−10 79 2687 -GM19631 LIB3056-007-Q1-N1-G9 LIB3056 g974781 BLASTN 388 1e−37 78 2688 -GM37189 LIB3051-072-Q1-K1-B5 LIB3051 g1814402 BLASTN 316 1e−15 78 2689 -GM44802 LIB3053-004-Q1-N1-B2 LIB3053 g974782 BLASTX 85 1e−26 90 2690 1382 700683236H1 SOYMON008 g886470 BLASTN 805 1e−58 78 2691 1382 700566434H1 SOYMON002 g886471 BLASTX 162 1e−23 76 2692 15690 701064361H1 SOYMON034 g974781 BLASTN 951 1e−70 84 2693 15690 700847301H1 SOYMON021 g886470 BLASTN 707 1e−66 85 2694 17335 LIB3051-088-Q1-K1-D9 LIB3051 g886470 BLASTN 1285 1e−101 79 2695 17335 701003832H1 SOYMON019 g974781 BLASTN 811 1e−58 77 2696 17335 700864888H1 SOYMON016 g1814402 BLASTN 767 1e−55 79 2697 17335 700672491H1 SOYMON006 g974781 BLASTN 428 1e−46 75 2698 17335 701003457H1 SOYMON019 g1814402 BLASTN 411 1e−29 84 2699 17335 700833672H1 SOYMON019 g974781 BLASTN 445 1e−28 79 2700 17900 700850578H1 SOYMON023 g2738247 BLASTN 905 1e−66 82 2701 17900 701053154H1 SOYMON032 g1814402 BLASTN 725 1e−65 83 2702 17900 700842351H1 SOYMON020 g1814402 BLASTN 878 1e−64 83 2703 17900 700837656H1 SOYMON020 g1814402 BLASTN 841 1e−61 83 2704 17900 700890851H1 SOYMON024 g974781 BLASTN 747 1e−53 80 2705 20688 700908840H1 SOYMON022 g2738247 BLASTN 889 1e−65 82 2706 20688 700908848H1 SOYMON022 g2738247 BLASTN 877 1e−64 81 2707 33542 LIB3051-009-Q1-E1-E5 LIB3051 g974781 BLASTN 1218 1e−92 79 2708 33542 700748773H1 SOYMON013 g974781 BLASTN 669 1e−46 78 2709 33542 700836363H1 SOYMON020 g2738247 BLASTN 596 1e−40 80 2710 4243 701123616H1 SOYMON037 g1814402 BLASTN 995 1e−74 87 2711 4243 700555001H1 SOYMON001 g1814402 BLASTN 975 1e−72 90 2712 4243 701002967H1 SOYMON019 g1814402 BLASTN 950 1e−70 86 2713 4243 700653509H1 SOYMON003 g1814402 BLASTN 539 1e−69 85 2714 4243 701206028H1 SOYMON035 g1814402 BLASTN 938 1e−69 86 2715 4243 700962115H1 SOYMON022 g1814402 BLASTN 923 1e−68 86 2716 4243 700866243H1 SOYMON016 g1814402 BLASTN 909 1e−66 82 2717 4243 700752507H1 SOYMON014 g1814402 BLASTN 887 1e−65 85 2718 4243 701003887H1 SOYMON019 g1814402 BLASTN 863 1e−63 86 2719 4243 700556913H1 SOYMON001 g1814402 BLASTN 865 1e−63 86 2720 4243 701013549H1 SOYMON019 g1814402 BLASTN 867 1e−63 90 2721 4243 701209706H1 SOYMON035 g1814402 BLASTN 871 1e−63 90 2722 4243 701010487H1 SOYMON019 g1814402 BLASTN 529 1e−62 79 2723 4243 700548246H1 SOYMON002 g1814402 BLASTN 553 1e−62 82 2724 4243 701138219H1 SOYMON038 g1814402 BLASTN 594 1e−62 86 2725 4243 700965160H1 SOYMON022 g1814402 BLASTN 852 1e−62 91 2726 4243 701015168H1 SOYMON019 g1814402 BLASTN 855 1e−62 89 2727 4243 701136095H1 SOYMON038 g1814402 BLASTN 839 1e−61 88 2728 4243 700761789H1 SOYMON015 g1814402 BLASTN 845 1e−61 86 2729 4243 701105695H1 SOYMON036 g1814402 BLASTN 835 1e−60 87 2730 4243 700991714H1 SOYMON011 g1814402 BLASTN 502 1e−59 83 2731 4243 700564223H1 SOYMON002 g1814402 BLASTN 557 1e−59 90 2732 4243 700987384H1 SOYMON009 g1814402 BLASTN 803 1e−58 86 2733 4243 700833934H1 SOYMON019 g1814402 BLASTN 806 1e−58 89 2734 4243 700835181H1 SOYMON019 g1814402 BLASTN 806 1e−58 82 2735 4243 700737529H1 SOYMON010 g1814402 BLASTN 810 1e−58 92 2736 4243 701012851H1 SOYMON019 g1814402 BLASTN 811 1e−58 91 2737 4243 700556592H1 SOYMON001 g1814402 BLASTN 814 1e−58 88 2738 4243 700907579H1 SOYMON022 g1814402 BLASTN 781 1e−56 89 2739 4243 700961749H1 SOYMON022 g1814402 BLASTN 785 1e−56 91 2740 4243 700835239H1 SOYMON019 g1814402 BLASTN 787 1e−56 86 2741 4243 700646425H1 SOYMON013 g1814402 BLASTN 772 1e−55 89 2742 4243 701123924H1 SOYMON037 g1814402 BLASTN 775 1e−55 91 2743 4243 700957759H1 SOYMON022 g1814402 BLASTN 776 1e−55 90 2744 4243 700964425H1 SOYMON022 g1814402 BLASTN 777 1e−55 86 2745 4243 700962173H1 SOYMON022 g1814402 BLASTN 778 1e−55 91 2746 4243 701066293H1 SOYMON034 g1814402 BLASTN 759 1e−54 81 2747 4243 700986741H1 SOYMON009 g1814402 BLASTN 589 1e−53 84 2748 4243 701212514H1 SOYMON035 g1814402 BLASTN 591 1e−53 87 2749 4243 701009195H1 SOYMON019 g1814402 BLASTN 747 1e−53 91 2750 4243 701060510H1 SOYMON033 g1814402 BLASTN 748 1e−53 84 2751 4243 700848730H1 SOYMON021 g1814402 BLASTN 749 1e−53 90 2752 4243 700754870H1 SOYMON014 g1814402 BLASTN 751 1e−53 84 2753 4243 701212482H1 SOYMON035 g1814402 BLASTN 751 1e−53 84 2754 4243 700753283H1 SOYMON014 g1814402 BLASTN 752 1e−53 86 2755 4243 700738517H1 SOYMON012 g1814402 BLASTN 636 1e−52 89 2756 4243 700833978H1 SOYMON019 g1814402 BLASTN 740 1e−52 91 2757 4243 700756424H1 SOYMON014 g1814402 BLASTN 729 1e−51 82 2758 4243 701011759H1 SOYMON019 g1814402 BLASTN 729 1e−51 91 2759 4243 701010103H2 SOYMON019 g1814402 BLASTN 707 1e−50 82 2760 4243 700741392H1 SOYMON012 g1814402 BLASTN 707 1e−50 84 2761 4243 701123062H1 SOYMON037 g1814402 BLASTN 308 1e−49 88 2762 4243 701048949H1 SOYMON032 g1814402 BLASTN 502 1e−49 85 2763 4243 700834566H1 SOYMON019 g1814402 BLASTN 618 1e−49 88 2764 4243 700963965H1 SOYMON022 g1814402 BLASTN 685 1e−48 78 2765 4243 700986376H1 SOYMON009 g1814402 BLASTN 694 1e−48 84 2766 4243 701012708H1 SOYMON019 g1814402 BLASTN 521 1e−47 91 2767 4243 700746927H1 SOYMON013 g1814402 BLASTN 547 1e−46 78 2768 4243 700997448H1 SOYMON018 g1814402 BLASTN 470 1e−45 89 2769 4243 700830667H1 SOYMON019 g1814402 BLASTN 647 1e−45 86 2770 4243 700891479H1 SOYMON024 g1814402 BLASTN 654 1e−45 88 2771 4243 700562246H1 SOYMON002 g1814402 BLASTN 656 1e−45 86 2772 4243 701097325H1 SOYMON028 g1814402 BLASTN 420 1e−44 92 2773 4243 700835830H1 SOYMON019 g1814402 BLASTN 503 1e−44 86 2774 4243 700962969H1 SOYMON022 g1814402 BLASTN 515 1e−44 89 2775 4243 701102860H1 SOYMON028 g1814402 BLASTN 309 1e−42 92 2776 4243 700763506H1 SOYMON015 g1814402 BLASTN 456 1e−42 84 2777 4243 700994059H1 SOYMON011 g1814402 BLASTN 487 1e−42 87 2778 4243 700751551H1 SOYMON014 g1814402 BLASTN 568 1e−41 85 2779 4243 701108872H1 SOYMON036 g1814402 BLASTN 603 1e−41 85 2780 4243 700994053H1 SOYMON011 g1814402 BLASTN 587 1e−40 85 2781 4243 700729064H1 SOYMON009 g1814402 BLASTN 413 1e−38 90 2782 4243 700874176H1 SOYMON018 g1814402 BLASTN 468 1e−38 92 2783 4243 700742963H1 SOYMON012 g1814402 BLASTN 565 1e−38 86 2784 4243 701212923H1 SOYMON035 g1814402 BLASTN 572 1e−38 84 2785 4243 700994851H1 SOYMON011 g1814402 BLASTN 548 1e−36 76 2786 4243 701000193H1 SOYMON018 g1814402 BLASTN 296 1e−33 87 2787 4243 700756219H1 SOYMON014 g1814402 BLASTN 394 1e−32 80 2788 4243 701014751H1 SOYMON019 g1814402 BLASTN 461 1e−29 90 2789 4243 700561163H1 SOYMON002 g1814402 BLASTN 231 1e−25 89 2790 4243 700650284H1 SOYMON003 g974782 BLASTX 157 1e−14 96 2791 4243 700869218H1 SOYMON016 g1814402 BLASTN 248 1e−11 93 2792 550 LIB3028-007-Q1-B1-B6 LIB3028 g886470 BLASTN 1440 1e−111 81 2793 550 LIB3040-017-Q1-E1-E8 LIB3040 g886470 BLASTN 1260 1e−96 80 2794 550 700650656H1 SOYMON003 g1814402 BLASTN 1072 1e−93 83 2795 550 LIB3051-091-Q1-K1-C2 LIB3051 g886470 BLASTN 1124 1e−89 80 2796 550 LIB3051-072-Q1-K1-B3 LIB3051 g974781 BLASTN 1114 1e−83 82 2797 550 LIB3051-006-Q1-E1-G9 LIB3051 g974781 BLASTN 703 1e−82 81 2798 550 700563811H1 SOYMON002 g1814402 BLASTN 1071 1e−80 84 2799 550 700754104H1 SOYMON014 g974781 BLASTN 1052 1e−78 86 2800 550 701002973H1 SOYMON019 g1814402 BLASTN 1039 1e−77 85 2801 550 700986733H1 SOYMON009 g974781 BLASTN 1039 1e−77 83 2802 550 700557595H1 SOYMON001 g886470 BLASTN 1019 1e−76 83 2803 550 700563477H1 SOYMON002 g2738247 BLASTN 1024 1e−76 85 2804 550 700976112H1 SOYMON009 g886470 BLASTN 1024 1e−76 84 2805 550 701004484H1 SOYMON019 g1814402 BLASTN 1007 1e−75 84 2806 550 700889161H1 SOYMON024 g974781 BLASTN 996 1e−74 85 2807 550 700833110H1 SOYMON019 g974781 BLASTN 998 1e−74 87 2808 550 701124559H1 SOYMON037 g974781 BLASTN 1002 1e−74 86 2809 550 700729207H1 SOYMON009 g974781 BLASTN 1004 1e−74 85 2810 550 700730012H1 SOYMON009 g886470 BLASTN 987 1e−73 85 2811 550 700987128H1 SOYMON009 g2738247 BLASTN 989 1e−73 85 2812 550 700564788H1 SOYMON002 g974781 BLASTN 990 1e−73 84 2813 550 701099104H1 SOYMON028 g1814402 BLASTN 994 1e−73 86 2814 550 LIB3065-008-Q1-N1-B4 LIB3065 g2738247 BLASTN 839 1e−72 83 2815 550 701104283H1 SOYMON036 g974781 BLASTN 976 1e−72 83 2816 550 700726387H1 SOYMON009 g974781 BLASTN 980 1e−72 84 2817 550 700900884H1 SOYMON027 g1814402 BLASTN 982 1e−72 85 2818 550 LIB3051-029-Q1-K1-D6 LIB3051 g886470 BLASTN 824 1e−71 81 2819 550 701213995H1 SOYMON035 g1814402 BLASTN 962 1e−71 84 2820 550 700683596H1 SOYMON008 g1814402 BLASTN 968 1e−71 85 2821 550 700646529H1 SOYMON014 g1814402 BLASTN 867 1e−70 84 2822 550 700756704H1 SOYMON014 g1814402 BLASTN 893 1e−70 87 2823 550 700991647H1 SOYMON011 g1814402 BLASTN 947 1e−70 84 2824 550 700962195H1 SOYMON022 g974781 BLASTN 947 1e−70 87 2825 550 700994369H1 SOYMON011 g1814402 BLASTN 949 1e−70 84 2826 550 700672477H1 SOYMON006 g1814402 BLASTN 951 1e−70 85 2827 550 700946215H1 SOYMON024 g2738247 BLASTN 957 1e−70 83 2828 550 701152765H1 SOYMON031 g974781 BLASTN 958 1e−70 88 2829 550 701010552H1 SOYMON019 g974781 BLASTN 501 1e−69 83 2830 550 LIB3056-004-Q1-N1-B12 LIB3056 g2738247 BLASTN 698 1e−69 77 2831 550 701100656H1 SOYMON028 g974781 BLASTN 890 1e−69 85 2832 550 700985362H1 SOYMON009 g1814402 BLASTN 937 1e−69 82 2833 550 700746178H1 SOYMON013 g974781 BLASTN 938 1e−69 85 2834 550 700674440H1 SOYMON007 g974781 BLASTN 938 1e−69 83 2835 550 700556934H1 SOYMON001 g1814402 BLASTN 938 1e−69 82 2836 550 700652624H1 SOYMON003 g1814402 BLASTN 939 1e−69 82 2837 550 700952331H1 SOYMON022 g886470 BLASTN 946 1e−69 83 2838 550 700725576H1 SOYMON009 g886470 BLASTN 946 1e−69 85 2839 550 701003602H1 SOYMON019 g2738247 BLASTN 924 1e−68 83 2840 550 700745053H1 SOYMON013 g974781 BLASTN 924 1e−68 85 2841 550 700895781H1 SOYMON027 g1814402 BLASTN 926 1e−68 84 2842 550 700664593H1 SOYMON005 g974781 BLASTN 926 1e−68 87 2843 550 700864264H1 SOYMON016 g1814402 BLASTN 930 1e−68 84 2844 550 700674466H1 SOYMON007 g974781 BLASTN 933 1e−68 83 2845 550 700564170H1 SOYMON002 g974781 BLASTN 861 1e−67 83 2846 550 700654531H1 SOYMON004 g974781 BLASTN 911 1e−67 81 2847 550 700996341H1 SOYMON018 g886470 BLASTN 911 1e−67 83 2848 550 701136257H1 SOYMON038 g886470 BLASTN 912 1e−67 82 2849 550 700751114H1 SOYMON014 g1814402 BLASTN 914 1e−67 84 2850 550 700657606H1 SOYMON004 g1814402 BLASTN 916 1e−67 87 2851 550 700983827H1 SOYMON009 g974781 BLASTN 918 1e−67 85 2852 550 700981249H1 SOYMON009 g1814402 BLASTN 921 1e−67 80 2853 550 700836103H1 SOYMON019 g886470 BLASTN 922 1e−67 82 2854 550 701011869H1 SOYMON019 g886470 BLASTN 486 1e−66 86 2855 550 701097045H1 SOYMON028 g886470 BLASTN 601 1e−66 85 2856 550 701012695H1 SOYMON019 g974781 BLASTN 777 1e−66 85 2857 550 700945396H1 SOYMON024 g974781 BLASTN 902 1e−66 85 2858 550 700755390H1 SOYMON014 g974781 BLASTN 903 1e−66 86 2859 550 700967721H1 SOYMON033 g886470 BLASTN 910 1e−66 82 2860 550 700750610H1 SOYMON014 g974781 BLASTN 910 1e−66 84 2861 550 700908231H1 SOYMON022 g974781 BLASTN 527 1e−65 83 2862 550 701003835H1 SOYMON019 g886470 BLASTN 723 1e−65 82 2863 550 700790802H1 SOYMON011 g2738247 BLASTN 891 1e−65 82 2864 550 701053438H1 SOYMON032 g974781 BLASTN 893 1e−65 82 2865 550 701009430H1 SOYMON019 g974781 BLASTN 894 1e−65 85 2866 550 700891415H1 SOYMON024 g1814402 BLASTN 895 1e−65 84 2867 550 701100681H1 SOYMON028 g2738247 BLASTN 482 1e−64 84 2868 550 700893420H1 SOYMON024 g974781 BLASTN 570 1e−64 89 2869 550 700752741H1 SOYMON014 g886470 BLASTN 880 1e−64 82 2870 550 700955938H1 SOYMON022 g1814402 BLASTN 882 1e−64 81 2871 550 701015042H1 SOYMON019 g2738247 BLASTN 886 1e−64 82 2872 550 LIB3051-072-Q1-K1-B1 LIB3051 g974781 BLASTN 645 1e−63 76 2873 550 700741918H1 SOYMON012 g886470 BLASTN 722 1e−63 84 2874 550 701103319H1 SOYMON028 g974781 BLASTN 792 1e−63 82 2875 550 700833218H1 SOYMON019 g2738247 BLASTN 863 1e−63 81 2876 550 701008071H1 SOYMON019 g886470 BLASTN 867 1e−63 83 2877 550 700832073H1 SOYMON019 g974781 BLASTN 871 1e−63 83 2878 550 700889695H1 SOYMON024 g974781 BLASTN 872 1e−63 83 2879 550 701007489H2 SOYMON019 g974781 BLASTN 873 1e−63 84 2880 550 700895858H1 SOYMON027 g974781 BLASTN 874 1e−63 84 2881 550 700753955H1 SOYMON014 g974781 BLASTN 470 1e−62 87 2882 550 700564433H1 SOYMON002 g886470 BLASTN 757 1e−62 84 2883 550 700963115H1 SOYMON022 g974781 BLASTN 851 1e−62 83 2884 550 700894728H1 SOYMON024 g886470 BLASTN 860 1e−62 84 2885 550 701056915H1 SOYMON033 g886470 BLASTN 862 1e−62 84 2886 550 700741134H1 SOYMON012 g886470 BLASTN 862 1e−62 83 2887 550 700847591H1 SOYMON021 g974781 BLASTN 842 1e−61 84 2888 550 700941253H1 SOYMON024 g2738247 BLASTN 844 1e−61 80 2889 550 701004315H1 SOYMON019 g2738247 BLASTN 846 1e−61 83 2890 550 700895720H1 SOYMON027 g974781 BLASTN 848 1e−61 83 2891 550 701013541H1 SOYMON019 g974781 BLASTN 849 1e−61 84 2892 550 700892552H1 SOYMON024 g886470 BLASTN 719 1e−60 83 2893 550 701141313H1 SOYMON038 g974781 BLASTN 827 1e−60 83 2894 550 701012547H1 SOYMON019 g1814402 BLASTN 831 1e−60 83 2895 550 701008558H1 SOYMON019 g1814402 BLASTN 831 1e−60 83 2896 550 700902022H1 SOYMON027 g2738247 BLASTN 831 1e−60 82 2897 550 700959515H1 SOYMON022 g886470 BLASTN 832 1e−60 81 2898 550 701042630H1 SOYMON029 g1814402 BLASTN 819 1e−59 81 2899 550 700941292H1 SOYMON024 g2738247 BLASTN 820 1e−59 81 2900 550 700788526H1 SOYMON011 g974781 BLASTN 491 1e−58 82 2901 550 700894839H1 SOYMON024 g1814402 BLASTN 498 1e−58 82 2902 550 700865873H1 SOYMON016 g974781 BLASTN 808 1e−58 85 2903 550 701015435H1 SOYMON019 g886470 BLASTN 809 1e−58 80 2904 550 700755960H1 SOYMON014 g2738247 BLASTN 809 1e−58 78 2905 550 700876051H1 SOYMON018 g886470 BLASTN 434 1e−57 81 2906 550 701041327H1 SOYMON029 g886470 BLASTN 499 1e−57 83 2907 550 701098902H1 SOYMON028 g2738247 BLASTN 767 1e−57 77 2908 550 700853392H1 SOYMON023 g886470 BLASTN 793 1e−57 82 2909 550 700872645H1 SOYMON018 g974781 BLASTN 798 1e−57 83 2910 550 700989675H1 SOYMON011 g2738247 BLASTN 800 1e−57 79 2911 550 700753487H1 SOYMON014 g1814402 BLASTN 589 1e−56 83 2912 550 700736276H1 SOYMON010 g1814402 BLASTN 782 1e−56 79 2913 550 700891361H1 SOYMON024 g1814402 BLASTN 783 1e−56 81 2914 550 700829712H1 SOYMON019 g974781 BLASTN 790 1e−56 80 2915 550 LIB3050-019-Q1-K1-A1 LIB3050 g974781 BLASTN 663 1e−55 81 2916 550 701001013H1 SOYMON018 g1814402 BLASTN 768 1e−55 84 2917 550 LIB3028-031-Q1-B1-G12 LIB3028 g886470 BLASTN 775 1e−55 82 2918 550 701212782H1 SOYMON035 g886470 BLASTN 621 1e−54 82 2919 550 701008695H1 SOYMON019 g974781 BLASTN 718 1e−54 80 2920 550 700990972H1 SOYMON011 g1814402 BLASTN 766 1e−54 81 2921 550 700789576H2 SOYMON011 g886470 BLASTN 766 1e−54 80 2922 550 700994266H1 SOYMON011 g1814402 BLASTN 408 1e−53 85 2923 550 700731985H1 SOYMON010 g974781 BLASTN 590 1e−53 81 2924 550 700907927H1 SOYMON022 g886470 BLASTN 612 1e−53 80 2925 550 701012079H1 SOYMON019 g974781 BLASTN 669 1e−53 86 2926 550 700753939H1 SOYMON014 g886470 BLASTN 690 1e−53 78 2927 550 701000754H1 SOYMON018 g974781 BLASTN 745 1e−53 76 2928 550 701040287H1 SOYMON029 g886470 BLASTN 747 1e−53 82 2929 550 700891329H1 SOYMON024 g974781 BLASTN 749 1e−53 79 2930 550 700897258H1 SOYMON027 g886470 BLASTN 752 1e−53 86 2931 550 700944949H1 SOYMON024 g974781 BLASTN 426 1e−52 82 2932 550 701108671H1 SOYMON036 g1814402 BLASTN 731 1e−52 77 2933 550 700905233H1 SOYMON022 g886470 BLASTN 732 1e−52 77 2934 550 700958589H1 SOYMON022 g886470 BLASTN 738 1e−52 80 2935 550 700666436H1 SOYMON005 g2738247 BLASTN 739 1e−52 82 2936 550 700829902H1 SOYMON019 g886470 BLASTN 740 1e−52 82 2937 550 700740110H1 SOYMON012 g974781 BLASTN 742 1e−52 83 2938 550 700989055H1 SOYMON011 g886470 BLASTN 538 1e−50 79 2939 550 701213370H1 SOYMON035 g974781 BLASTN 599 1e−50 83 2940 550 701098072H1 SOYMON028 g974781 BLASTN 709 1e−50 74 2941 550 700896128H1 SOYMON027 g886470 BLASTN 714 1e−50 83 2942 550 701060755H1 SOYMON033 g974781 BLASTN 716 1e−50 74 2943 550 700953594H1 SOYMON022 g1814402 BLASTN 695 1e−49 78 2944 550 701046911H1 SOYMON032 g2738247 BLASTN 687 1e−48 81 2945 550 701065707H1 SOYMON034 g1814402 BLASTN 443 1e−47 84 2946 550 700962114H1 SOYMON022 g886470 BLASTN 678 1e−47 84 2947 550 700831826H1 SOYMON019 g2738247 BLASTN 682 1e−47 77 2948 550 701054296H1 SOYMON032 g886470 BLASTN 454 1e−46 83 2949 550 700888738H1 SOYMON024 g974781 BLASTN 552 1e−46 77 2950 550 700892022H1 SOYMON024 g886470 BLASTN 605 1e−46 81 2951 550 700890275H1 SOYMON024 g2738247 BLASTN 666 1e−46 77 2952 550 LIB3051-006-Q1-K1-G9 LIB3051 g974781 BLASTN 670 1e−45 80 2953 550 700889113H1 SOYMON024 g886470 BLASTN 582 1e−43 81 2954 550 700952720H1 SOYMON022 g886470 BLASTN 611 1e−42 76 2955 550 701014761H1 SOYMON019 g886470 BLASTN 318 1e−41 84 2956 550 700753882H1 SOYMON014 g886470 BLASTN 381 1e−41 79 2957 550 700743792H1 SOYMON012 g1814402 BLASTN 610 1e−41 86 2958 550 700990963H1 SOYMON011 g886470 BLASTN 578 1e−39 77 2959 550 700941880H1 SOYMON024 g2738247 BLASTN 583 1e−39 81 2960 550 700898962H1 SOYMON027 g2738247 BLASTN 571 1e−38 80 2961 550 700990865H1 SOYMON011 g2738247 BLASTN 467 1e−37 77 2962 550 700565779H1 SOYMON002 g886470 BLASTN 557 1e−37 70 2963 550 700993903H1 SOYMON011 g974781 BLASTN 562 1e−37 84 2964 550 700941589H1 SOYMON024 g2738247 BLASTN 550 1e−36 81 2965 550 701052554H1 SOYMON032 g886470 BLASTN 534 1e−35 67 2966 550 700991055H1 SOYMON011 g886470 BLASTN 356 1e−34 79 2967 550 701010438H1 SOYMON019 g2738247 BLASTN 508 1e−33 79 2968 550 700756634H1 SOYMON014 g2738247 BLASTN 494 1e−32 76 2969 550 701042980H1 SOYMON029 g886470 BLASTN 461 1e−29 83 2970 550 700682940H1 SOYMON008 g2738247 BLASTN 466 1e−29 83 2971 550 701049575H1 SOYMON032 g886470 BLASTN 433 1e−27 82 2972 550 700982552H1 SOYMON009 g886470 BLASTN 356 1e−25 78 2973 550 700675637H1 SOYMON007 g886470 BLASTN 375 1e−25 79 2974 550 701142153H1 SOYMON038 g886470 BLASTN 377 1e−22 76 2975 550 700682724H1 SOYMON008 g974781 BLASTN 361 1e−19 88 2976 550 701051764H1 SOYMON032 g1814402 BLASTN 211 1e−17 80 2977 550 700867241H1 SOYMON016 g2738248 BLASTX 152 1e−13 88 2978 550 701054954H1 SOYMON032 g2738248 BLASTX 138 1e−12 86 2979 550 700790450H2 SOYMON011 g974781 BLASTN 238 1e−10 80 2980 550 700653979H1 SOYMON003 g1814403 BLASTX 118 1e−9 92 2981 550 700894218H1 SOYMON024 g2738248 BLASTX 122 1e−9 78 2982 550 700863078H1 SOYMON022 g2738247 BLASTN 236 1e−8 78 2983 5758 701209304H1 SOYMON035 g886470 BLASTN 766 1e−54 84 2984 5758 701106455H1 SOYMON036 g886470 BLASTN 723 1e−51 82 2985 5758 700833538H1 SOYMON019 g1814402 BLASTN 609 1e−43 83 2986 5758 701051425H1 SOYMON032 g886470 BLASTN 629 1e−43 83 2987 5758 700654506H1 SOYMON004 g886470 BLASTN 438 1e−26 70 2988 5758 701047795H1 SOYMON032 g974782 BLASTX 161 1e−15 100 2989 5758 701202409H1 SOYMON035 g1814402 BLASTN 310 1e−15 79 2990 8266 700558628H1 SOYMON001 g886470 BLASTN 780 1e−56 74 2991 8266 701207720H1 SOYMON035 g1814402 BLASTN 766 1e−54 74 2992 8266 700557429H1 SOYMON001 g1814402 BLASTN 728 1e−51 75

ADENOSYLHOMOCYSTEINASE (EC 3.3.1.1) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 1354 -700154280 700154280H1 SATMON007 g170772 BLASTN 229 1e−23 77 1355 -L30594291 LIB3059-032-Q1-K1-A8 LIB3059 g170772 BLASTN 516 1e−74 73 1356 -L30664307 LIB3066-047-Q1-K1-E7 LIB3066 g170772 BLASTN 628 1e−49 70 1357 503 LIB3079-013-Q1-K1-D3 LIB3079 g170772 BLASTN 1505 1e−139 89 1358 503 LIB3062-017-Q1-K1-A10 LIB3062 g170772 BLASTN 1571 1e−129 89 1359 503 LIB3067-001-Q1-K1-D11 LIB3067 g170772 BLASTN 1654 1e−129 89 1360 503 LIB148-060-Q1-E1-B9 LIB148 g170772 BLASTN 1620 1e−126 90 1361 503 LIB3069-043-Q1-K1-G4 LIB3069 g170772 BLASTN 1360 1e−124 84 1362 503 LIB143-006-Q1-E1-F3 LIB143 g170772 BLASTN 831 1e−120 88 1363 503 LIB189-009-Q1-E1-E9 LIB189 g170772 BLASTN 1551 1e−120 90 1364 503 LIB3060-003-Q1-K1-F9 LIB3060 g170772 BLASTN 1538 1e−119 91 1365 503 700089023H1 SATMON011 g170772 BLASTN 1495 1e−115 91 1366 503 LIB143-061-Q1-E1-C9 LIB143 g170772 BLASTN 1474 1e−114 90 1367 503 LIB3069-034-Q1-K1-E5 LIB3069 g170772 BLASTN 1483 1e−114 88 1368 503 LIB143-061-Q1-E1-E5 LIB143 g170772 BLASTN 1440 1e−113 89 1369 503 LIB3067-040-Q1-K1-H5 LIB3067 g170772 BLASTN 1467 1e−113 88 1370 503 LIB3067-048-Q1-K1-A11 LIB3067 g170772 BLASTN 1207 1e−109 86 1371 503 LIB3068-050-Q1-K1-G6 LIB3068 g170772 BLASTN 1279 1e−109 86 1372 503 LIB3069-036-Q1-K1-F6 LIB3069 g170772 BLASTN 1310 1e−109 92 1373 503 LIB3066-047-Q1-K1-H5 LIB3066 g170772 BLASTN 1353 1e−108 86 1374 503 LIB3059-011-Q1-K1-A5 LIB3059 g170772 BLASTN 1401 1e−107 90 1375 503 LIB3067-056-Q1-K1-E11 LIB3067 g170772 BLASTN 957 1e−106 86 1376 503 LIB189-010-Q1-E1-E12 LIB189 g170772 BLASTN 1144 1e−106 88 1377 503 700084426H1 SATMON011 g170772 BLASTN 1368 1e−105 91 1378 503 700086273H1 SATMON011 g170772 BLASTN 1364 1e−104 91 1379 503 700573027H1 SATMON030 g170772 BLASTN 939 1e−103 89 1380 503 700209360H1 SATMON016 g170772 BLASTN 1350 1e−103 89 1381 503 700619916H1 SATMON034 g170772 BLASTN 1050 1e−102 89 1382 503 700086051H1 SATMON011 g170772 BLASTN 1336 1e−102 90 1383 503 LIB3069-034-Q1-K1-C8 LIB3069 g170772 BLASTN 1322 1e−101 86 1384 503 700026324H1 SATMON003 g170772 BLASTN 1328 1e−101 91 1385 503 700104549H1 SATMON010 g170772 BLASTN 836 1e−100 88 1386 503 700622108H1 SATMON034 g170772 BLASTN 1158 1e−100 88 1387 503 700093980H1 SATMON008 g170772 BLASTN 1312 1e−100 90 1388 503 700077427H1 SATMON007 g170772 BLASTN 1317 1e−100 90 1389 503 700095389H1 SATMON008 g170772 BLASTN 1302 1e−99 91 1390 503 LIB3067-055-Q1-K1-D3 LIB3067 g170772 BLASTN 762 1e−98 87 1391 503 LIB3060-054-Q1-K1-F6 LIB3060 g170772 BLASTN 1009 1e−98 83 1392 503 700083339H1 SATMON011 g170772 BLASTN 1282 1e−98 91 1393 503 700102631H1 SATMON010 g170772 BLASTN 1283 1e−98 89 1394 503 700265625H1 SATMON017 g170772 BLASTN 1286 1e−98 90 1395 503 700095002H1 SATMON008 g170772 BLASTN 1289 1e−98 88 1396 503 700094761H1 SATMON008 g170772 BLASTN 1289 1e−98 88 1397 503 700073832H1 SATMON007 g170772 BLASTN 1270 1e−97 88 1398 503 700047817H1 SATMON003 g170772 BLASTN 1272 1e−97 91 1399 503 700091149H1 SATMON011 g170772 BLASTN 1262 1e−96 89 1400 503 700098584H1 SATMON009 g170772 BLASTN 1264 1e−96 91 1401 503 700085932H1 SATMON011 g170772 BLASTN 1266 1e−96 89 1402 503 700094081H1 SATMON008 g170772 BLASTN 1269 1e−96 91 1403 503 700202442H1 SATMON003 g170772 BLASTN 1145 1e−95 88 1404 503 700049770H1 SATMON003 g170772 BLASTN 1246 1e−95 89 1405 503 LIB3079-020-Q1-K1-B12 LIB3079 g170772 BLASTN 1249 1e−95 85 1406 503 700090226H1 SATMON011 g170772 BLASTN 1252 1e−95 90 1407 503 700088191H1 SATMON011 g170772 BLASTN 1253 1e−95 90 1408 503 700076743H1 SATMON007 g170772 BLASTN 1256 1e−95 90 1409 503 LIB189-009-Q1-E1-E10 LIB189 g170772 BLASTN 789 1e−94 88 1410 503 700072038H1 SATMON007 g170772 BLASTN 1237 1e−94 89 1411 503 700082967H1 SATMON011 g170772 BLASTN 1239 1e−94 89 1412 503 LIB3068-062-Q1-K1-A3 LIB3068 g170772 BLASTN 1148 1e−93 82 1413 503 700094713H1 SATMON008 g170772 BLASTN 1222 1e−93 88 1414 503 700095620H1 SATMON008 g170772 BLASTN 1173 1e−92 89 1415 503 700242509H1 SATMON010 g170772 BLASTN 1213 1e−92 92 1416 503 700071923H1 SATMON007 g170772 BLASTN 1214 1e−92 90 1417 503 700575314H1 SATMON030 g170772 BLASTN 1149 1e−91 88 1418 503 700086654H1 SATMON011 g170772 BLASTN 1199 1e−91 90 1419 503 700241072H1 SATMON010 g170772 BLASTN 1205 1e−91 91 1420 503 700047361H1 SATMON003 g170772 BLASTN 1110 1e−90 89 1421 503 700217056H1 SATMON016 g170772 BLASTN 1187 1e−90 91 1422 503 LIB3067-005-Q1-K1-F2 LIB3067 g170772 BLASTN 1188 1e−90 84 1423 503 700075495H1 SATMON007 g170772 BLASTN 1193 1e−90 86 1424 503 700084783H1 SATMON011 g170772 BLASTN 1194 1e−90 91 1425 503 LIB143-059-Q1-E1-C3 LIB143 g170772 BLASTN 952 1e−89 84 1426 503 700448818H1 SATMON028 g170772 BLASTN 1174 1e−89 91 1427 503 700159079H1 SATMON012 g170772 BLASTN 1177 1e−89 92 1428 503 700094408H1 SATMON008 g170772 BLASTN 1177 1e−89 92 1429 503 700348440H1 SATMON023 g170772 BLASTN 1182 1e−89 89 1430 503 700077082H1 SATMON007 g170772 BLASTN 1182 1e−89 92 1431 503 700082111H1 SATMON011 g170772 BLASTN 1183 1e−89 89 1432 503 700239752H1 SATMON010 g170772 BLASTN 1164 1e−88 89 1433 503 700029456H1 SATMON003 g170772 BLASTN 1166 1e−88 90 1434 503 700209430H1 SATMON016 g170772 BLASTN 1170 1e−88 90 1435 503 700208660H1 SATMON016 g170772 BLASTN 930 1e−87 90 1436 503 700213138H1 SATMON016 g170772 BLASTN 1042 1e−87 89 1437 503 700102234H1 SATMON010 g170772 BLASTN 634 1e−85 90 1438 503 700450479H1 SATMON028 g170772 BLASTN 1015 1e−85 90 1439 503 700095372H1 SATMON008 g170772 BLASTN 1126 1e−85 90 1440 503 700218734H1 SATMON011 g170772 BLASTN 1128 1e−85 94 1441 503 700256858H1 SATMON017 g170772 BLASTN 1133 1e−85 90 1442 503 700221063H1 SATMON011 g170772 BLASTN 1137 1e−85 90 1443 503 700105439H1 SATMON010 g170772 BLASTN 1118 1e−84 89 1444 503 700085174H1 SATMON011 g170772 BLASTN 1125 1e−84 90 1445 503 700242421H1 SATMON010 g170772 BLASTN 1125 1e−84 92 1446 503 700077492H1 SATMON007 g170772 BLASTN 774 1e−83 88 1447 503 700209295H1 SATMON016 g170772 BLASTN 914 1e−83 89 1448 503 700455826H1 SATMON029 g170772 BLASTN 978 1e−83 91 1449 503 700213370H1 SATMON016 g170772 BLASTN 1110 1e−83 92 1450 503 700352095H1 SATMON023 g170772 BLASTN 1111 1e−83 89 1451 503 700076842H1 SATMON007 g170772 BLASTN 1112 1e−83 90 1452 503 700215872H1 SATMON016 g170772 BLASTN 1113 1e−83 89 1453 503 700073645H1 SATMON007 g170772 BLASTN 1113 1e−83 89 1454 503 700048153H1 SATMON003 g170772 BLASTN 651 1e−82 90 1455 503 700073307H1 SATMON007 g170772 BLASTN 762 1e−82 90 1456 503 700350009H1 SATMON023 g170772 BLASTN 924 1e−82 88 1457 503 LIB143-059-Q1-E1-C5 LIB143 g170772 BLASTN 1029 1e−82 83 1458 503 700073955H1 SATMON007 g170772 BLASTN 1090 1e−82 90 1459 503 LIB3059-042-Q1-K1-H12 LIB3059 g170772 BLASTN 1090 1e−82 85 1460 503 700238024H1 SATMON010 g170772 BLASTN 1091 1e−82 90 1461 503 700155863H1 SATMON007 g170772 BLASTN 1091 1e−82 93 1462 503 700217890H1 SATMON016 g170772 BLASTN 1093 1e−82 91 1463 503 700239314H1 SATMON010 g170772 BLASTN 1094 1e−82 88 1464 503 700048337H1 SATMON003 g170772 BLASTN 1094 1e−82 92 1465 503 700235469H1 SATMON010 g170772 BLASTN 1098 1e−82 91 1466 503 700164224H1 SATMON013 g170772 BLASTN 710 1e−81 92 1467 503 700209374H1 SATMON016 g170772 BLASTN 1083 1e−81 89 1468 503 700082268H1 SATMON011 g170772 BLASTN 1083 1e−81 88 1469 503 700159326H1 SATMON012 g170772 BLASTN 777 1e−80 91 1470 503 700025934H1 SATMON003 g170772 BLASTN 792 1e−80 91 1471 503 700210458H1 SATMON016 g170772 BLASTN 856 1e−80 88 1472 503 700243826H1 SATMON010 g170772 BLASTN 881 1e−80 90 1473 503 700242676H1 SATMON010 g170772 BLASTN 940 1e−80 89 1474 503 700345565H1 SATMON021 g170772 BLASTN 1034 1e−80 89 1475 503 700264733H1 SATMON017 g170772 BLASTN 1044 1e−80 90 1476 503 700159161H1 SATMON012 g170772 BLASTN 1068 1e−80 90 1477 503 700072491H1 SATMON007 g170772 BLASTN 604 1e−79 88 1478 503 700053204H1 SATMON008 g170772 BLASTN 618 1e−79 89 1479 503 700451515H1 SATMON028 g170772 BLASTN 1057 1e−79 85 1480 503 700468929H1 SATMON025 g170772 BLASTN 1061 1e−79 84 1481 503 700205801H1 SATMON003 g170772 BLASTN 1064 1e−79 91 1482 503 700611326H1 SATMON022 g170772 BLASTN 725 1e−78 88 1483 503 700082291H1 SATMON011 g170772 BLASTN 1043 1e−78 87 1484 503 700071695H1 SATMON007 g170772 BLASTN 1048 1e−78 89 1485 503 700551561H1 SATMON022 g170772 BLASTN 746 1e−77 88 1486 503 700221019H1 SATMON011 g170772 BLASTN 856 1e−77 89 1487 503 LIB3078-007-Q1-K1-C5 LIB3078 g170772 BLASTN 936 1e−77 83 1488 503 700049925H1 SATMON003 g170772 BLASTN 954 1e−77 89 1489 503 700154101H1 SATMON007 g170772 BLASTN 1036 1e−77 88 1490 503 700380151H1 SATMON021 g170772 BLASTN 613 1e−76 88 1491 503 700243831H1 SATMON010 g170772 BLASTN 842 1e−76 89 1492 503 700235671H1 SATMON010 g170772 BLASTN 1019 1e−76 90 1493 503 700087847H1 SATMON011 g170772 BLASTN 1009 1e−75 82 1494 503 700208747H1 SATMON016 g170772 BLASTN 1010 1e−75 89 1495 503 700071795H1 SATMON007 g170772 BLASTN 1011 1e−75 89 1496 503 700157002H1 SATMON012 g170772 BLASTN 1012 1e−75 90 1497 503 700201608H1 SATMON003 g170772 BLASTN 1015 1e−75 85 1498 503 700451541H1 SATMON028 g170772 BLASTN 1015 1e−75 86 1499 503 700212182H1 SATMON016 g170772 BLASTN 1015 1e−75 88 1500 503 700381485H1 SATMON023 g170772 BLASTN 1017 1e−75 91 1501 503 700096793H1 SATMON008 g170772 BLASTN 761 1e−74 87 1502 503 700093025H1 SATMON008 g170772 BLASTN 996 1e−74 89 1503 503 700017129H1 SATMON001 g170772 BLASTN 998 1e−74 92 1504 503 700216525H1 SATMON016 g170772 BLASTN 1000 1e−74 86 1505 503 700087912H1 SATMON011 g170772 BLASTN 1001 1e−74 91 1506 503 700172618H1 SATMON013 g170772 BLASTN 1001 1e−74 91 1507 503 700801894H1 SATMON036 g170772 BLASTN 1002 1e−74 90 1508 503 700162040H1 SATMON012 g170772 BLASTN 1003 1e−74 92 1509 503 700212084H1 SATMON016 g170772 BLASTN 1004 1e−74 89 1510 503 700027971H1 SATMON003 g170772 BLASTN 982 1e−73 92 1511 503 700077321H1 SATMON007 g170772 BLASTN 985 1e−73 89 1512 503 700619884H1 SATMON034 g170772 BLASTN 987 1e−73 87 1513 503 700457201H1 SATMON029 g170772 BLASTN 988 1e−73 87 1514 503 700082970H1 SATMON011 g170772 BLASTN 990 1e−73 89 1515 503 700083350H1 SATMON011 g170772 BLASTN 971 1e−72 89 1516 503 700017732H1 SATMON001 g170772 BLASTN 973 1e−72 88 1517 503 700094688H1 SATMON008 g170772 BLASTN 975 1e−72 89 1518 503 700170985H1 SATMON013 g170772 BLASTN 979 1e−72 89 1519 503 700451847H1 SATMON028 g170772 BLASTN 872 1e−71 84 1520 503 700455091H1 SATMON029 g170772 BLASTN 931 1e−71 93 1521 503 700106189H1 SATMON010 g170772 BLASTN 960 1e−71 88 1522 503 700160075H1 SATMON012 g170772 BLASTN 968 1e−71 90 1523 503 700084952H1 SATMON011 g170772 BLASTN 552 1e−70 78 1524 503 700348238H1 SATMON023 g170772 BLASTN 946 1e−70 87 1525 503 700151310H1 SATMON007 g170772 BLASTN 948 1e−70 90 1526 503 700048962H1 SATMON003 g170772 BLASTN 677 1e−69 88 1527 503 700088485H1 SATMON011 g170772 BLASTN 936 1e−69 88 1528 503 700093667H1 SATMON008 g170772 BLASTN 937 1e−69 88 1529 503 700161491H1 SATMON012 g170772 BLASTN 928 1e−68 86 1530 503 700073939H1 SATMON007 g170772 BLASTN 931 1e−68 88 1531 503 700027715H1 SATMON003 g170772 BLASTN 472 1e−67 90 1532 503 700072843H1 SATMON007 g170772 BLASTN 692 1e−67 87 1533 503 700439356H1 SATMON026 g170772 BLASTN 777 1e−67 85 1534 503 700073556H1 SATMON007 g170772 BLASTN 910 1e−67 88 1535 503 700154461H1 SATMON007 g170772 BLASTN 911 1e−67 87 1536 503 700201909H1 SATMON003 g170772 BLASTN 912 1e−67 88 1537 503 700104384H1 SATMON010 g170772 BLASTN 665 1e−66 89 1538 503 700152707H1 SATMON007 g170772 BLASTN 898 1e−66 88 1539 503 700076625H1 SATMON007 g170772 BLASTN 900 1e−66 85 1540 503 700456945H1 SATMON029 g170772 BLASTN 902 1e−66 80 1541 503 700457294H1 SATMON029 g170772 BLASTN 446 1e−65 88 1542 503 700152062H1 SATMON007 g170772 BLASTN 803 1e−65 90 1543 503 700072428H2 SATMON007 g170772 BLASTN 890 1e−65 88 1544 503 700217352H1 SATMON016 g170772 BLASTN 893 1e−65 88 1545 503 700072443H2 SATMON007 g170772 BLASTN 894 1e−65 89 1546 503 700087436H1 SATMON011 g170772 BLASTN 874 1e−64 88 1547 503 700240615H1 SATMON010 g170772 BLASTN 877 1e−64 88 1548 503 700155233H1 SATMON007 g170772 BLASTN 883 1e−64 90 1549 503 700241210H1 SATMON010 g170772 BLASTN 822 1e−63 87 1550 503 700478070H1 SATMON025 g170772 BLASTN 863 1e−63 82 1551 503 700447433H1 SATMON027 g170772 BLASTN 865 1e−63 84 1552 503 700215884H1 SATMON016 g170772 BLASTN 865 1e−63 88 1553 503 700575250H1 SATMON030 g170772 BLASTN 820 1e−61 84 1554 503 700209283H1 SATMON016 g170772 BLASTN 833 1e−60 86 1555 503 700213479H1 SATMON016 g170772 BLASTN 834 1e−60 88 1556 503 700241740H1 SATMON010 g170772 BLASTN 814 1e−59 88 1557 503 700026012H1 SATMON003 g170772 BLASTN 815 1e−59 87 1558 503 700151036H1 SATMON007 g170772 BLASTN 816 1e−59 91 1559 503 700207150H1 SATMON017 g170772 BLASTN 818 1e−59 87 1560 503 700217383H1 SATMON016 g170772 BLASTN 824 1e−59 88 1561 503 700618168H1 SATMON033 g170772 BLASTN 385 1e−58 80 1562 503 700020777H1 SATMON001 g170772 BLASTN 806 1e−58 85 1563 503 700224131H1 SATMON011 g170772 BLASTN 794 1e−57 88 1564 503 700222876H1 SATMON011 g170772 BLASTN 794 1e−57 88 1565 503 700027716H1 SATMON003 g170772 BLASTN 642 1e−56 79 1566 503 700073305H1 SATMON007 g170772 BLASTN 787 1e−56 83 1567 503 700257376H1 SATMON017 g170772 BLASTN 362 1e−55 87 1568 503 700151752H1 SATMON007 g170772 BLASTN 766 1e−55 89 1569 503 700219086H1 SATMON011 g170772 BLASTN 769 1e−55 87 1570 503 700218924H1 SATMON011 g170772 BLASTN 775 1e−55 87 1571 503 700424527H1 SATMONN01 g170772 BLASTN 777 1e−55 83 1572 503 700073183H1 SATMON007 g170772 BLASTN 607 1e−54 85 1573 503 700217412H1 SATMON016 g170772 BLASTN 755 1e−54 87 1574 503 700208634H1 SATMON016 g170772 BLASTN 642 1e−53 83 1575 503 700202577H1 SATMON003 g170772 BLASTN 743 1e−53 86 1576 503 700446645H1 SATMON027 g170772 BLASTN 744 1e−53 87 1577 503 700238061H1 SATMON010 g170772 BLASTN 745 1e−53 86 1578 503 700617325H1 SATMON033 g170772 BLASTN 751 1e−53 91 1579 503 700239901H1 SATMON010 g170772 BLASTN 641 1e−52 86 1580 503 700155130H1 SATMON007 g170772 BLASTN 646 1e−52 88 1581 503 700083360H1 SATMON011 g170772 BLASTN 739 1e−52 87 1582 503 700236915H1 SATMON010 g170772 BLASTN 740 1e−52 87 1583 503 700074780H1 SATMON007 g170772 BLASTN 528 1e−51 83 1584 503 700479515H1 SATMON034 g170772 BLASTN 718 1e−51 88 1585 503 700215545H1 SATMON016 g170772 BLASTN 724 1e−51 90 1586 503 700165354H1 SATMON013 g170772 BLASTN 728 1e−51 86 1587 503 700353994H1 SATMON024 g170772 BLASTN 713 1e−50 87 1588 503 700153619H1 SATMON007 g170772 BLASTN 694 1e−49 86 1589 503 700074779H1 SATMON007 g170772 BLASTN 705 1e−49 77 1590 503 700155530H1 SATMON007 g170772 BLASTN 686 1e−48 86 1591 503 700221207H1 SATMON011 g170772 BLASTN 692 1e−48 86 1592 503 700264257H1 SATMON017 g170772 BLASTN 672 1e−47 87 1593 503 700152216H1 SATMON007 g170772 BLASTN 659 1e−46 88 1594 503 700150643H1 SATMON007 g170772 BLASTN 659 1e−46 88 1595 503 700156027H1 SATMON007 g170772 BLASTN 655 1e−45 93 1596 503 700260335H1 SATMON017 g170772 BLASTN 385 1e−44 79 1597 503 700623795H1 SATMON034 g170772 BLASTN 401 1e−44 88 1598 503 700150581H1 SATMON007 g170772 BLASTN 640 1e−44 88 1599 503 700151970H1 SATMON007 g170772 BLASTN 645 1e−44 87 1600 503 700151780H1 SATMON007 g170772 BLASTN 628 1e−43 88 1601 503 700575547H1 SATMON030 g170772 BLASTN 614 1e−42 90 1602 503 LIB143-026-Q1-E1-A5 LIB143 g170772 BLASTN 633 1e−42 77 1603 503 700347945H1 SATMON023 g170772 BLASTN 586 1e−40 84 1604 503 700074416H1 SATMON007 g170772 BLASTN 589 1e−40 86 1605 503 700352990H1 SATMON024 g170772 BLASTN 575 1e−39 92 1606 503 700156025H1 SATMON007 g170772 BLASTN 568 1e−38 91 1607 503 700432072H1 SATMONN01 g170772 BLASTN 374 1e−37 84 1608 503 700354249H1 SATMON024 g170772 BLASTN 528 1e−35 93 1609 503 700617977H1 SATMON033 g170772 BLASTN 535 1e−35 87 1610 503 700218312H1 SATMON016 g170772 BLASTN 236 1e−33 91 1611 503 700456080H1 SATMON029 g170772 BLASTN 513 1e−33 83 1612 503 700349395H1 SATMON023 g170772 BLASTN 500 1e−32 91 1613 503 700051637H1 SATMON003 g170772 BLASTN 249 1e−31 86 1614 503 700202153H1 SATMON003 g170772 BLASTN 474 1e−30 91 1615 503 700256951H1 SATMON017 g170772 BLASTN 454 1e−29 86 1616 503 700150542H1 SATMON007 g170772 BLASTN 343 1e−28 76 1617 503 700151629H1 SATMON007 g170772 BLASTN 445 1e−28 85 1618 503 700446654H1 SATMON027 g170772 BLASTN 437 1e−27 84 1619 503 700155814H1 SATMON007 g170772 BLASTN 428 1e−26 88 1620 503 700161019H1 SATMON012 g2588780 BLASTN 417 1e−25 92 1621 503 700377236H1 SATMON019 g170772 BLASTN 402 1e−24 84 1622 503 LIB3067-036-Q1-K1-A4 LIB3067 g1220121 BLASTN 224 1e−22 84 1623 503 700158933H1 SATMON012 g170772 BLASTN 368 1e−21 90 1624 503 700155365H1 SATMON007 g170772 BLASTN 347 1e−20 91 1625 503 700405366H1 SATMON029 g170772 BLASTN 311 1e−17 88 1626 503 700159812H1 SATMON012 g407412 BLASTX 160 1e−15 96 1627 503 700209759H1 SATMON016 g170772 BLASTN 239 1e−15 90 1628 503 700154527H1 SATMON007 g170772 BLASTN 250 1e−12 92 1629 503 700449637H1 SATMON028 g170772 BLASTN 201 1e−10 78 1630 503 700096829H1 SATMON008 g170772 BLASTN 216 1e−9 89 2993 -700661285 700661285H1 SOYMON005 g1857024 BLASTX 95 1e−12 100 2994 -700750570 700750570H1 SOYMON014 g170772 BLASTN 414 1e−24 81 2995 -700752735 700752735H1 SOYMON014 g170772 BLASTN 446 1e−27 78 2996 -700755052 700755052H1 SOYMON014 g170772 BLASTN 547 1e−45 74 2997 -700756501 700756501H1 SOYMON014 g535583 BLASTN 717 1e−50 84 2998 -700831127 700831127H1 SOYMON019 g535583 BLASTN 862 1e−63 83 2999 -700851779 700851779H1 SOYMON023 g170772 BLASTN 505 1e−33 77 3000 -700888715 700888715H1 SOYMON024 g535583 BLASTN 442 1e−31 91 3001 -700889420 700889420H1 SOYMON024 g1220121 BLASTN 893 1e−65 84 3002 -700895218 700895218H1 SOYMON024 g407411 BLASTN 816 1e−59 83 3003 -700941379 700941379H1 SOYMON024 g170772 BLASTN 424 1e−33 72 3004 -700986855 700986855H1 SOYMON009 g170772 BLASTN 701 1e−49 74 3005 -701070484 701070484H1 SOYMON034 g407411 BLASTN 362 1e−43 73 3006 -701136279 701136279H1 SOYMON038 g170772 BLASTN 651 1e−56 80 3007 -GM16478 LIB3054-007-Q1-N1-G3 LIB3054 g2244750 BLASTX 70 1e−27 61 3008 -GM23819 LIB3040-019-Q1-E1-C5 LIB3040 g535583 BLASTN 258 1e−10 88 3009 -GM29758 LIB3050-016-Q1-E1-B11 LIB3050 g535583 BLASTN 391 1e−42 70 3010 16 LIB3030-003-Q1-B1-B11 LIB3030 g3088578 BLASTN 1577 1e−122 87 3011 16 LIB3050-023-Q1-K1-H9 LIB3050 g535583 BLASTN 1498 1e−116 86 3012 16 LIB3030-003-Q1-B1-F7 LIB3030 g535583 BLASTN 1469 1e−113 86 3013 16 LIB3055-005-Q1-N1-C11 LIB3055 g170772 BLASTN 1205 1e−109 84 3014 16 LIB3065-011-Q1-N1-A3 LIB3065 g170772 BLASTN 622 1e−107 88 3015 16 700652256H1 SOYMON003 g170772 BLASTN 591 1e−90 88 3016 16 LIB3065-011-Q1-N1-A4 LIB3065 g170772 BLASTN 1182 1e−89 78 3017 16 700653827H1 SOYMON003 g170772 BLASTN 660 1e−87 81 3018 16 701099940H1 SOYMON028 g1220121 BLASTN 1154 1e−87 88 3019 16 701003671H1 SOYMON019 g170772 BLASTN 1131 1e−85 92 3020 16 700752105H1 SOYMON014 g170772 BLASTN 1116 1e−84 91 3021 16 700653057H1 SOYMON003 g170772 BLASTN 990 1e−83 88 3022 16 700945531H1 SOYMON024 g535583 BLASTN 1109 1e−83 89 3023 16 700980013H1 SOYMON009 g535583 BLASTN 1109 1e−83 88 3024 16 701127812H1 SOYMON037 g170772 BLASTN 1110 1e−83 91 3025 16 LIB3056-014-Q1-N1-F8 LIB3056 g170772 BLASTN 798 1e−81 82 3026 16 700653862H1 SOYMON003 g1220121 BLASTN 960 1e−80 88 3027 16 700994148H1 SOYMON011 g535583 BLASTN 1069 1e−80 86 3028 16 700984184H1 SOYMON009 g170772 BLASTN 756 1e−79 86 3029 16 701123715H1 SOYMON037 g1220121 BLASTN 1059 1e−79 87 3030 16 700839038H1 SOYMON020 g170772 BLASTN 1060 1e−79 93 3031 16 700978445H1 SOYMON009 g170772 BLASTN 1062 1e−79 89 3032 16 701123035H1 SOYMON037 g170772 BLASTN 829 1e−78 90 3033 16 701041545H1 SOYMON029 g170772 BLASTN 1030 1e−77 86 3034 16 700898192H1 SOYMON027 g1220121 BLASTN 1031 1e−77 89 3035 16 700985750H1 SOYMON009 g169662 BLASTN 1033 1e−77 85 3036 16 700730995H1 SOYMON009 g170772 BLASTN 1040 1e−77 91 3037 16 700555909H1 SOYMON001 g535583 BLASTN 593 1e−76 84 3038 16 700746538H1 SOYMON013 g535583 BLASTN 834 1e−76 88 3039 16 700941380H1 SOYMON024 g170772 BLASTN 991 1e−76 91 3040 16 701118851H1 SOYMON037 g170772 BLASTN 1019 1e−76 89 3041 16 701065379H1 SOYMON034 g535583 BLASTN 1022 1e−76 86 3042 16 701209645H1 SOYMON035 g170772 BLASTN 795 1e−75 86 3043 16 701055017H1 SOYMON032 g1220121 BLASTN 877 1e−75 86 3044 16 701015213H1 SOYMON019 g535583 BLASTN 1008 1e−75 87 3045 16 700982770H1 SOYMON009 g1220121 BLASTN 1009 1e−75 85 3046 16 701212420H1 SOYMON035 g535583 BLASTN 1012 1e−75 86 3047 16 700977916H1 SOYMON009 g170772 BLASTN 700 1e−74 89 3048 16 700974401H1 SOYMON005 g1220121 BLASTN 857 1e−74 89 3049 16 700645749H1 SOYMON010 g170772 BLASTN 996 1e−74 82 3050 16 700646620H1 SOYMON014 g170772 BLASTN 996 1e−74 89 3051 16 701126103H1 SOYMON037 g170772 BLASTN 1000 1e−74 88 3052 16 700978001H1 SOYMON009 g170772 BLASTN 563 1e−73 89 3053 16 700561987H1 SOYMON002 g1220121 BLASTN 782 1e−73 87 3054 16 701101526H1 SOYMON028 g1220121 BLASTN 810 1e−73 88 3055 16 700562680H1 SOYMON002 g170772 BLASTN 985 1e−73 87 3056 16 700560521H1 SOYMON001 g170772 BLASTN 988 1e−73 80 3057 16 701055544H1 SOYMON032 g170772 BLASTN 992 1e−73 88 3058 16 701061418H1 SOYMON033 g170772 BLASTN 993 1e−73 87 3059 16 701049514H1 SOYMON032 g1220121 BLASTN 884 1e−72 88 3060 16 700646215H1 SOYMON012 g170772 BLASTN 971 1e−72 89 3061 16 701014875H1 SOYMON019 g535583 BLASTN 973 1e−72 87 3062 16 700874718H1 SOYMON018 g169662 BLASTN 974 1e−72 86 3063 16 700897796H1 SOYMON027 g1220121 BLASTN 977 1e−72 87 3064 16 700548019H1 SOYMON001 g170772 BLASTN 547 1e−71 87 3065 16 700904875H1 SOYMON022 g535583 BLASTN 963 1e−71 88 3066 16 701106315H1 SOYMON036 g170772 BLASTN 557 1e−70 90 3067 16 700745023H1 SOYMON013 g407411 BLASTN 949 1e−70 85 3068 16 701120348H1 SOYMON037 g170772 BLASTN 956 1e−70 86 3069 16 701124508H1 SOYMON037 g170772 BLASTN 578 1e−69 84 3070 16 700956183H1 SOYMON022 g1220121 BLASTN 625 1e−69 87 3071 16 700894619H1 SOYMON024 g535583 BLASTN 828 1e−69 85 3072 16 700892392H1 SOYMON024 g535583 BLASTN 936 1e−69 85 3073 16 700730676H1 SOYMON009 g535583 BLASTN 939 1e−69 86 3074 16 700728913H1 SOYMON009 g407411 BLASTN 940 1e−69 87 3075 16 700845737H1 SOYMON021 g1220121 BLASTN 528 1e−68 88 3076 16 700990961H1 SOYMON011 g170772 BLASTN 582 1e−68 85 3077 16 700983443H1 SOYMON009 g170772 BLASTN 922 1e−68 86 3078 16 701120754H1 SOYMON037 g170772 BLASTN 923 1e−68 88 3079 16 700900486H1 SOYMON027 g1220121 BLASTN 924 1e−68 83 3080 16 701133574H2 SOYMON038 g170772 BLASTN 929 1e−68 88 3081 16 700900924H1 SOYMON027 g170772 BLASTN 931 1e−68 88 3082 16 701056706H1 SOYMON032 g170772 BLASTN 486 1e−67 89 3083 16 701110051H1 SOYMON036 g170772 BLASTN 910 1e−67 88 3084 16 701136325H1 SOYMON038 g170772 BLASTN 915 1e−67 87 3085 16 700750809H1 SOYMON014 g170772 BLASTN 900 1e−66 88 3086 16 700686607H1 SOYMON008 g170772 BLASTN 901 1e−66 88 3087 16 700848261H1 SOYMON021 g535583 BLASTN 905 1e−66 87 3088 16 700686634H1 SOYMON008 g170772 BLASTN 905 1e−66 88 3089 16 700891285H1 SOYMON024 g170772 BLASTN 906 1e−66 88 3090 16 700560291H1 SOYMON001 g170772 BLASTN 906 1e−66 88 3091 16 700752975H1 SOYMON014 g170772 BLASTN 907 1e−66 89 3092 16 701006013H2 SOYMON019 g170772 BLASTN 908 1e−66 87 3093 16 700974038H1 SOYMON005 g1220121 BLASTN 631 1e−65 88 3094 16 701047024H1 SOYMON032 g170772 BLASTN 707 1e−65 89 3095 16 700900409H1 SOYMON027 g170772 BLASTN 737 1e−65 83 3096 16 701137320H1 SOYMON038 g170772 BLASTN 769 1e−65 88 3097 16 700978805H1 SOYMON009 g170772 BLASTN 886 1e−65 84 3098 16 700726195H1 SOYMON009 g1220121 BLASTN 889 1e−65 87 3099 16 700661112H1 SOYMON005 g170772 BLASTN 661 1e−64 85 3100 16 700989712H1 SOYMON011 g170772 BLASTN 788 1e−64 88 3101 16 700752287H1 SOYMON014 g170772 BLASTN 875 1e−64 85 3102 16 700964226H1 SOYMON022 g170772 BLASTN 877 1e−64 88 3103 16 700847346H1 SOYMON021 g170772 BLASTN 880 1e−64 83 3104 16 700756428H1 SOYMON014 g170772 BLASTN 883 1e−64 88 3105 16 701049928H1 SOYMON032 g170772 BLASTN 885 1e−64 82 3106 16 700848652H1 SOYMON021 g170772 BLASTN 477 1e−63 89 3107 16 700898929H1 SOYMON027 g1220121 BLASTN 514 1e−63 87 3108 16 700903523H1 SOYMON022 g170772 BLASTN 527 1e−63 82 3109 16 700983745H1 SOYMON009 g1220121 BLASTN 863 1e−63 88 3110 16 700890587H1 SOYMON024 g170772 BLASTN 865 1e−63 86 3111 16 700969917H1 SOYMON005 g407411 BLASTN 868 1e−63 83 3112 16 700808487H1 SOYMON024 g170772 BLASTN 870 1e−63 88 3113 16 700749968H1 SOYMON013 g170772 BLASTN 871 1e−63 87 3114 16 700751254H1 SOYMON014 g170772 BLASTN 575 1e−62 87 3115 16 701014277H1 SOYMON019 g170772 BLASTN 739 1e−62 86 3116 16 700853635H1 SOYMON023 g170772 BLASTN 854 1e−62 87 3117 16 700752357H1 SOYMON014 g170772 BLASTN 856 1e−62 88 3118 16 700754523H1 SOYMON014 g170772 BLASTN 856 1e−62 92 3119 16 700982153H1 SOYMON009 g170772 BLASTN 400 1e−61 82 3120 16 700958283H1 SOYMON022 g170772 BLASTN 839 1e−61 87 3121 16 700980911H1 SOYMON009 g170772 BLASTN 842 1e−61 82 3122 16 700788112H1 SOYMON011 g170772 BLASTN 844 1e−61 83 3123 16 701005927H1 SOYMON019 g170772 BLASTN 849 1e−61 88 3124 16 700756443H1 SOYMON014 g170772 BLASTN 849 1e−61 88 3125 16 700658914H1 SOYMON004 g1220121 BLASTN 468 1e−60 85 3126 16 701135266H1 SOYMON038 g170772 BLASTN 827 1e−60 87 3127 16 700982179H1 SOYMON009 g170772 BLASTN 827 1e−60 82 3128 16 700754593H1 SOYMON014 g170772 BLASTN 832 1e−60 81 3129 16 700831723H1 SOYMON019 g170772 BLASTN 814 1e−59 91 3130 16 700986775H1 SOYMON009 g170772 BLASTN 815 1e−59 92 3131 16 700755219H1 SOYMON014 g170772 BLASTN 820 1e−59 84 3132 16 701015494H1 SOYMON019 g170772 BLASTN 822 1e−59 88 3133 16 701008473H1 SOYMON019 g170772 BLASTN 823 1e−59 87 3134 16 700754981H1 SOYMON014 g170772 BLASTN 824 1e−59 88 3135 16 700870790H1 SOYMON018 g170772 BLASTN 825 1e−59 81 3136 16 700833069H1 SOYMON019 g170772 BLASTN 806 1e−58 86 3137 16 700680127H2 SOYMON008 g535583 BLASTN 807 1e−58 86 3138 16 701015374H1 SOYMON019 g170772 BLASTN 807 1e−58 87 3139 16 700872895H1 SOYMON018 g535583 BLASTN 808 1e−58 88 3140 16 701137912H1 SOYMON038 g170772 BLASTN 374 1e−57 83 3141 16 700984063H1 SOYMON009 g1220121 BLASTN 575 1e−57 79 3142 16 700991988H1 SOYMON011 g170772 BLASTN 791 1e−57 82 3143 16 700873915H1 SOYMON018 g170772 BLASTN 793 1e−57 88 3144 16 700978721H1 SOYMON009 g170772 BLASTN 797 1e−57 78 3145 16 701213679H1 SOYMON035 g170772 BLASTN 798 1e−57 88 3146 16 701102954H1 SOYMON028 g170772 BLASTN 799 1e−57 79 3147 16 700888289H1 SOYMON024 g170772 BLASTN 712 1e−56 91 3148 16 700962086H1 SOYMON022 g170772 BLASTN 783 1e−56 88 3149 16 701052227H1 SOYMON032 g535583 BLASTN 788 1e−56 86 3150 16 700755177H1 SOYMON014 g170772 BLASTN 768 1e−55 88 3151 16 701123136H1 SOYMON037 g170772 BLASTN 771 1e−55 82 3152 16 700979067H1 SOYMON009 g170772 BLASTN 776 1e−55 90 3153 16 700755513H1 SOYMON014 g170772 BLASTN 759 1e−54 92 3154 16 700756639H1 SOYMON014 g170772 BLASTN 761 1e−54 81 3155 16 700554077H1 SOYMON001 g170772 BLASTN 371 1e−53 86 3156 16 700653194H1 SOYMON003 g170772 BLASTN 395 1e−53 86 3157 16 701110348H1 SOYMON036 g170772 BLASTN 743 1e−53 81 3158 16 700753973H1 SOYMON014 g170772 BLASTN 743 1e−53 87 3159 16 701011009H1 SOYMON019 g535583 BLASTN 733 1e−52 81 3160 16 700739086H1 SOYMON012 g170772 BLASTN 456 1e−51 87 3161 16 700740140H1 SOYMON012 g170772 BLASTN 723 1e−51 89 3162 16 700565040H1 SOYMON002 g170772 BLASTN 729 1e−51 74 3163 16 701148186H1 SOYMON031 g535583 BLASTN 665 1e−50 85 3164 16 701142734H1 SOYMON038 g535583 BLASTN 670 1e−47 86 3165 16 700754441H1 SOYMON014 g170772 BLASTN 385 1e−46 93 3166 16 701102588H1 SOYMON028 g1220121 BLASTN 662 1e−46 89 3167 16 700900656H1 SOYMON027 g535583 BLASTN 635 1e−44 87 3168 16 700974207H1 SOYMON005 g535583 BLASTN 641 1e−44 85 3169 16 700982081H1 SOYMON009 g170772 BLASTN 494 1e−43 78 3170 16 701130026H1 SOYMON037 g1220121 BLASTN 482 1e−42 86 3171 16 701009720H1 SOYMON019 g170772 BLASTN 621 1e−42 87 3172 16 700962602H1 SOYMON022 g170772 BLASTN 357 1e−40 91 3173 16 700724934H1 SOYMON009 g535583 BLASTN 561 1e−40 81 3174 16 700729305H1 SOYMON009 g535583 BLASTN 544 1e−39 82 3175 16 701210054H1 SOYMON035 g535583 BLASTN 569 1e−38 85 3176 16 700790192H1 SOYMON011 g535583 BLASTN 293 1e−37 83 3177 16 700984076H1 SOYMON009 g170772 BLASTN 198 1e−35 88 3178 16 700726562H1 SOYMON009 g170772 BLASTN 307 1e−34 78 3179 16 701211376H1 SOYMON035 g170772 BLASTN 524 1e−34 86 3180 16 700753085H1 SOYMON014 g2588780 BLASTN 358 1e−33 76 3181 16 700727993H1 SOYMON009 g535583 BLASTN 465 1e−32 84 3182 16 700561072H1 SOYMON001 g170772 BLASTN 473 1e−30 83 3183 16 701211464H1 SOYMON035 g170772 BLASTN 464 1e−28 81 3184 16 701098045H1 SOYMON028 g169660 BLASTN 386 1e−21 78 3185 16 700945233H1 SOYMON024 g407412 BLASTX 150 1e−18 87 3186 16 700752655H1 SOYMON014 g758247 BLASTX 172 1e−16 94 3187 16 700735356H1 SOYMON010 g758247 BLASTX 152 1e−14 100 3188 16 700683995H1 SOYMON008 g758247 BLASTX 106 1e−13 90 3189 16 700658760H1 SOYMON004 g1857024 BLASTX 123 1e−13 63 3190 16 700762885H1 SOYMON015 g1857024 BLASTX 134 1e−13 89 3191 16 700755740H1 SOYMON014 g170773 BLASTX 149 1e−13 100 3192 16 700854969H1 SOYMON023 g170772 BLASTN 178 1e−12 80 3193 16 701143036H1 SOYMON038 g169661 BLASTX 113 1e−8 83 3194 18409 700786561H1 SOYMON011 g535583 BLASTN 992 1e−73 85 3195 18409 701008057H1 SOYMON019 g535583 BLASTN 831 1e−60 87 3196 18409 701037442H1 SOYMON029 g535583 BLASTN 669 1e−46 86 3197 18409 700942865H1 SOYMON024 g535583 BLASTN 464 1e−32 86 3198 7322 700651524H1 SOYMON003 g170772 BLASTN 466 1e−65 80 3199 7322 700565758H1 SOYMON002 g170772 BLASTN 450 1e−63 81

CYSTATHIONINE β-SYNTHASE (EC 4.2.1.22) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 1631 −700025795 700025795H1 SATMON003 g1323263 BLASTX 186 1e−25 68 1632 20651 700344783H1 SATMON021 g1813975 BLASTX 41 1e−9 53

CYSTATHIONINE γ-LYASE (EC 4.4.1.1) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 1633 −700260027 700260027H1 SATMON017 g169475 BLASTX 112 1e−10 75 1634 1228 700027629H1 SATMON003 g169475 BLASTX 189 1e−19 87 3203 −700750583 700750583H1 SOYMON014 g169475 BLASTX 149 1e−13 78 3204 12502 LIB3051-069-Q1-K1-E6 LIB3051 g2641242 BLASTX 86 1e−30 38

O-ACETYLHOMOSERINE (THIOL)-LYASE (EC 4.2.99.10) Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 3200 12502 701135185H1 SOYMON038 g1628606 BLASTX 100 1e−10 48 3201 12502 701042913H1 SOYMON029 g2605905 BLASTX 110 1e−9 42 3202 12502 701059330H1 SOYMON033 g2605905 BLASTX 110 1e−9 42 *Table Headings Cluster ID

A cluster ID is arbitrarily assigned to all of those clones which belong to the same cluster at a given stringency and a particular clone will belong to only one cluster at a given stringency. If a cluster contains only a single clone (a “singleton”), then the cluster ID number will be negative, with an absolute value equal to the clone ID number of its single member. The cluster ID entries in the table refer to the cluster with which the particular clone in each row is associated.

Clone ID

The clone ID number refers to the particular clone in the PhytoSeq database. Each clone ID entry in the table refers to the clone whose sequence is used for (1) the sequence comparison whose scores are presented and/or (2) assignment to the particular cluster which is presented. Note that a clone may be included in this table even if its sequence comparison scores fail to meet the minimum standards for similarity. In such a case, the clone is included due solely to its association with a particular cluster for which sequences of one or more other member clones possess the required level of similarity.

Library

The library ID refers to the particular cDNA library from which a given clone is obtained. Each cDNA library is associated with the particular tissue(s), line(s) and developmental stage(s) from which it is isolated.

NCBI gi

Each sequence in the GENBANK public database is arbitrarily assigned a unique NCBI gi (National Center for Biotechnology Information GenBank Identifier) number. In this table, the NCBI gi number which is associated (in the same row) with a given clone refers to the particular GenBank sequence which is used in the sequence comparison. This entry is omitted when a clone is included solely due to its association with a particular cluster.

Method

The entry in the “Method” column of the table refers to the type of BLAST search that is used for the sequence comparison. “CLUSTER” is entered when the sequence comparison scores for a given clone fail to meet the minimum values required for significant similarity. In such cases, the clone is listed in the table solely as a result of its association with a given cluster for which sequences of one or more other member clones possess the required level of similarity.

Score

Each entry in the “Score” column of the table refers to the BLAST score that is generated by sequence comparison of the designated clone with the designated GENBANK sequence using the designated BLAST method. This entry is omitted when a clone is included solely due to its association with a particular cluster. If the program used to determine the hit is HMMSW then the score refers to HMMSW score.

P-Value

The entries in the P-Value column refer to the probability that such matches occur by chance.

%Ident

The entries in the “%Ident” column of the table refer to the percentage of identically matched nucleotides (or residues) that exist along the length of that portion of the sequences which is aligned by the BLAST comparison to generate the statistical scores presented. This entry is omitted when a clone is included solely due to its association with a particular cluster. 

1. A substantially purified nucleic acid molecule that encodes a maize S-adenosyl-methionine decarboxylase fragment wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with SEQ ID NO:
 662. 2. A substantially purified nucleic acid molecule comprising a nucleic acid sequence which shares between 100% and 95% sequence identity with SEQ ID NO: 662 or complete complement thereof.
 3. The substantially purified nucleic acid molecule of claim 2, wherein said substantially purified nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 98% sequence identity with SEQ ID NO: 662 or complete complement thereof.
 4. The substantially purified nucleic acid molecule of claim 2, wherein said substantially purified nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 99% sequence identity with SEQ ID NO: 662 or complete complement thereof.
 5. The substantially purified nucleic acid molecule of claim 2, wherein said substantially purified nucleic acid molecule comprises a nucleic acid sequence with 100% sequence identity with SEQ ID NO: 662 or complete complement thereof.
 6. A transformed plant comprising a nucleic acid molecule which comprises: (a) an exogenous promoter region which functions in a cell of the plant to cause the production of an mRNA molecule, which is linked to, (b) a structural nucleic acid molecule, wherein said structural mucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with SEQ ID NO: 622 or complete complement thereof, which is linked to (c) a 3′ non-translated sequence that functions in the plant cell to cause the termination of transcription and addition of polyadenylated ribonucleotides to the 3′ end of the mRNA molecule.
 7. The transformed plant of claim 6, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 98% sequence identity with SEQ ID NO: 662 or complete complement thereof.
 8. The transformed plant of claim 6, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 99% sequence identity with SEQ ID NO: 662 or complete complement thereof.
 9. The transformed plant of claim 6, wherein said nucleic acid molecule comprises a nucleic acid sequence with 100% sequence identity with SEQ ID NO: 662 or complete complement thereof.
 10. A transformed seed comprising a transformed plant cell comprising a nucleic acid molecule which comprises: (a) an exogenous promoter region which functions in said plant cell to cause the production of an mRNA molecule; which is linked to; (b) a structural nucleic acid molecule, wherein said structural nucleic acid molecule comprises a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 95% sequence identity with SEQ ID NO: 662 or complete complement thereof, which is linked to (c) a 3′ non-translated sequence that functions in said plant cell to cause the termination of transcription and the addition of polyadenylated ribonucleotides to said 3′ end of said mRNA molecule.
 11. The transformed seed according to claim 10, wherein said nucleic acid sequence is the complement of a nucleic acid sequence of SEQ ID NO:
 662. 12. The transformed seed according to claim 10, wherein said exogenous promoter region functions in a seed cell.
 13. The transformed seed according to claim 10, wherein said nucleic acid sequence shares between 100% and 98% sequence identity with SEQ ID NO: 662 or complete complement thereof.
 14. The transformed seed according to claim 10, wherein said nucleic acid sequence shares between 100% and 99% sequence identity with SEQ ID NO: 662 or complete complement thereof.
 15. The transformed seed according to claim 10, wherein said nucleic acid sequence shares 100% sequence identity with SEQ ID NO: 662 or complete complement thereof. 